Antiglaucoma agents

ABSTRACT

Pharmaceutical compositions and a method for reducing intraocular pressure by topically applying a carboxyalkyl dipeptide are disclosed.

This is a division of application Ser. No. 892,003, filed July 30, 1986,which will issue as U.S. Pat. No. 4,885,293 and which is acontinuation-in-part of Ser. No. 784,000 filed Oct. 4, 1985, now U.S.Pat. No. 4,826,816 and Ser. No. 721,015, filed Apr. 8, 1985, now U.S.Pat. No. 4,634,698, each of which are a continuation-in-part of Ser. No.653,186, filed Sept. 24, 1984, now U.S. Pat. No. 4,556,655.

The present invention relates to ophthalmological methods andpharmaceutical compositions employing carboxyalkyl dipeptides joinedthrough a sulfonamido group to a benzothiadiazinyl or sulfonylphenylmoiety.

BACKGROUND OF THE INVENTION

Glaucoma is an ocular disease complex associated with an elevatedpressure within the eye (i.e., intraocular pressure, IOP). As a resultof the elevated IOP, damage to the optic nerve head resulting inirreversible loss of visual function may ensue. Untreated, thiscondition may eventually lead to blindness.

Ocular hypertension, i.e., the condition of elevated intraocularpressure without optic nerve head damage or characteristic glaucomatousvisual field loss, is now believed by the majority of ophthalmologiststo represent the earliest phase in the onset of glaucoma.

A number of the drugs presently employed to treat glaucoma are notentirely satisfactory, particularly in the earliest course of thedisease when the side effects they produce are often worse than thesymptoms of the disease.

Epinephrine, used as a topical solution, must be utilized cautiously inpatients with high blood pressure, diabetes, hyperthyroidism andcerebral artereosclerosis due to the possibility of systemic action.

Timolol, a clinically utilized, topically applied agent for loweringintraocular pressure, must be used with caution in patients in whombeta-adrenergic blockade may be undesirable. Systemic absorption oftopically administered timolol and systemic beta-blockade areresponsible for the contraindication of timolol therapy for glaucoma inpatients with compromised pulmonary function and in patients who cannottolerate its systemic cardiovascular action.

Pilocarpine, a topical drug, although considered systemically harmlessand quite effective, may cause considerable local difficulties. Pupilconstriction causes the eye to lose its ability to adapt from light todark. Accommodation may become stimulated so that the patient'srefraction is sometimes incorrect and vision becomes blurred. The drugitself may cause a local vasodilation and red eyes. Irritation iscommon.

Carbonic anhydrase inhibitors have been used systemically but they havea number of disadvantages. While effective in lowering intraocularpressure, they often cause a numbness and tingling, gastrointestinalupsets and, frequently, depression, lethargy, a loss of appetite, andcentral malaise. European Patent Application 81400326.5, Publicationnumber 36,351 attempts to overcome these difficulties by the topicaladministration of an alkali metal salt of a carbonic anhydraseinhibitor.

The present invention provides a new composition and method for reducingand controlling elevated intraocular pressure, especially the elevatedIOP associated with glaucoma.

SUMMARY OF THE INVENTION

The invention in a pharmaceutical composition aspect involves a topicalophthalmologically acceptable composition useful for reducing andcontrollinq elevated intraocular pressure, especially elevated IOPassociated with glaucoma, which comprises an intraocular pressurereducing effective amount of a compound of formula I as described belowin combination with an ophthalmologically acceptable carrier for topicaluse. The invention in a pharmaceutical method aspect involves a methodfor controlling elevated intraocular pressure in an eye, especiallyelevated IOP associated with glaucoma in a human eye, which methodcomprises administering to said eye an intraocular pressure reducingeffective amount of a compound of formula I. Such method and compositionmay also be used in conjunction with the administration of abeta-adrenergic blocking agent and/or an anti-inflammatory steroid.

The compounds contemplated for use in the compositions and methods ofthis invention have the following structural formula I: ##STR1## or apharmaceutically acceptable salt thereof, wherein: A is ##STR2## k is 1or 2; n is 0 or 1;

p and q are 0, 1 or 2, provided that in structures IIb and IIc the sumof p and q is 1 or 2, and that in formula IId, p is not 0;

B is --[J]--[L]--[M]--;

D is ##STR3## E is --NH--, --O--, --S--, or --CH₂ --; G is ##STR4##--SO₂ --; J is --(CH₂)_(s) -- or --((CH₂)_(t) --W)--;

L is a chemical bond, cis- or trans-lower alkenylene, lower alkynylene,-Z-aryl-, -aryl-Z-, -Z-cycloalkyl-, or -cycloalkyl-Z-, a 5- or6-membered heterocyclic radical comprising 3 to 5 carbon atoms and 1 or2 heteroatoms selected from N, O and S, or a R⁵ -substitutedheterocyclic radical, wherein aryl is ##STR5## and cycloalkyl is##STR6## wherein w is 1, 2 or 3:

M is --(CH₂)_(u) -- or --((CH₂)_(t) --X--(CH₂)_(v))--;

W is ##STR7## X and Z are independently a chemical bond, --NR⁹ --,--O--, --S--, ##STR8## s, u and v are independently 0-5; t is 1-5;

R¹, R² and R⁹ are independently hydrogen, lower alkyl or lower acyl;

R³ is hydrogen, lower alkyl, halo- and dihaloloweralkyl,trifluoroethylthiomethyl, phenylloweralkyl, (cycloalkyl)lower-alkyl,aminomethyl, lower alkylaminomethyl,phenylloweralkylaminomethyl,(cycloalkyl)loweralkylaminomethyl,loweralkylthiomethyl, haloloweralkylthiomethyl, 2-,3- or4-pyridylloweralkyl, 2-, 4- or 5-thiazolylloweralkyl, 2-, 4- or5-1H-imidazolylloweralkyl, 1-imidazolylloweralkyl,1-morpholinoloweralkyl or hydroxyphenylloweralkyl;

R⁴ is chlorine or CF₃ ;

R⁵ is hydrogen, halogen, lower alkyl, lower acyl, lower alkoxy,haloloweralkyl or phenylloweralkyl;

R⁷ is hydrogen, lower alkyl or aminoloweralkyl;

R⁶ and R⁸ are independently hydroxy, alkoxy having from 1 to 8 carbonatoms, benzyl, allyl, R¹⁰ --Q_(r) --(CH₂)_(m) --O--, wherein Q is oxygenor sulfur, r is 0 or 1 and m is 2 to 4, ##STR9## wherein the alkyl hasfrom 3 to 8 carbon atoms, ##STR10## wherein the phenyl may besubstituted with group T defined, below, 1-glyceryl, ##STR11## R¹⁰ isphenyl, substituted phenyl wherein the substituents are chosen fromgroup T, 1-naphthyl or 2-naphthyl;

T is halogen, hydroxy, trifluoromethyl, lower alkoxy, lower alkyl,2-furanyl, 3-furanyl, 2-thienyl, 3-thienyl, phenyl and substitutedphenyl wherein the substituents are chosen from halogen, hydroxy,trifluoromethyl, lower alkoxy or lower alkyl;

R¹¹ is hydrogen or alkyl having from 1 to 8 carbon atoms;

R¹² is hydrogen, lower alkyl, unsubstituted or substituted phenyl andsubstituted or unsubstituted phenyl lower alkyl wherein phenyl may besubstituted by group T; and

R¹³ is hydrogen or lower alkyl;

provided that if L is alkenylene or alkynylene, J is --(CH₂)_(s) --wherein s is 1-5; provided that if L is -Z-aryl- or -Z-cycloalkyl-, J is--(CH₂)_(s) -- wherein s is 2-5; provided that if L is alkenylene,alkynylene, -aryl-Z- or -cycloalkyl-Z-, M is --(CH₂)_(u) -- wherein u is1-5; provided that if s and u are each zero, L is aryl or cycloalkyl(i.e. Z is a bond); and provided that if s and v are each zero, L isaryl or cycloalkyl (i.e. Z is a bond); and

with the further provisos that, when D is of formula IIIb and R¹ ishydrogen, B is not --(CH₂)₄ --; and that, when D is of formula IIIb andR¹ is hydrogen or lower alkyl, B is not --(CH₂)_(s) --aryl--(CH₂)_(t)--X--(CH₂)_(v) -- wherein s is 0 or 1, t is 1, v is 0 to 2 and X is abond, --O--, or --S--.

The invention in another aspect involves two kits for use in reducingand controlling elevated intraocular pressure. Both kits comprise firstand second containers, in a single package, wherein in both cases thefirst container includes a topical pharmaceutical composition comprisingan IOP reducing effective amount of a compound of formula I. In a firstkit, the second container includes a pharmaceutical compositioncomprising an anti-inflammatory effective amount of a steroid in apharmaceutically acceptable carrier. In the second kit, the secondcontainer includes a pharmaceutical composition comprising anintraocular pressure reducing amount of a beta adrenergic blocking agentin a topical opthamological carrier.

DETAILED DESCRIPTION

The compounds employed in the method and composition of the presentinvention are the subject matter of Ser. No. 721,015, filed Apr. 8,1985; Ser. No. 784,000, filed Oct. 4, 1985; and Ser. No. 653,186, filedSept. 24, 1984, which issued as U.S. Pat. No. 4,556,655, the disclosuresof which are incorporated herein by reference.

When A is formula IIb or IIc, the preferred sum of p and q is 1; when Ais of formula IId, preferred values for each of p and q are 1. Compoundswherein A is lIa or IIb wherein p is 0 and o is 1, and IId wherein p andq are each 1 and n is zero are preferred.

Also preferred are compounds wherein D is of formula IIIa, withcompounds wherein R⁴ is chlorine and G is --SO₂ -- being more preferred.Another group of preferred compounds are those wherein D is of formulaIIIa, R⁴ is chlorine, G is --SO₂ --, R² is preferably hydrogen and R³ ispreferably phenylethyl, (cyclopentyl)methyl, chloromethyl,dichloromethyl, 2-pyridinylethyl, butyl, pentyl, ortrifluoroethylthiomethyl.

Also preferred are compounds wherein B is ##STR12## i.e., compoundswherein J is --(CH₂)_(s) -- and s is 0-1, L is a bond or -Z-aryl-wherein Z is a bond, and M is --((CH₂)_(t) --X--(CH₂)_(v)), wherein t is1 , v is 1-2, and X is a bond or --O--.

For compounds wherein L is a heterocyclic radical, thiazolyl ringsjoined to the rest of molecule at the 2,4 and 2,5 positions arepreferred.

A preferred group for E is --NH--.

Another group of preferred compounds are those wherein R⁷ is hydrogen,methyl or aminobutyl. Still another group of preferred compounds arethose wherein R⁶ and R⁸ are hydroxy, ethoxy, methoxy, phenoxyethoxy,1-glyceryl, pivaloyloxyalkyloxy, and ##STR13##

A preferred group of compounds are those represented by the generalformula ##STR14## wherein R³ is phenylethyl, chloromethyl,dichloromethyl, butyl, pentyl, (cyclopentyl)methyl,trifluoroethylthiomethyl or 2-pyridinylethyl.

As used herein, "lower alkyl" means straight or branched chainhydrocarbon radicals of from 1 to 6 carbons, e.g. methyl, ethyl, propyl,isopropyl, butyl, t-butyl, pentyl and hexyl. Similarly, "lower alkoxy"means straight or branched alkoxy radicals having from 1 to 6 carbonatoms, e.g. methoxy, ethoxy, propoxy, butoxy, iso-butoxy, pentoxy andhexyloxy. "Halogen" means fluorine, chlorine and bromine. "Lower acyl"means organic radicals obtained by removing the hydroxyl group from thecorresponding carboxylic acid of from 1 to 6 carbons, e.g. formyl,acetyl, propionyl and butyryl. "Lower alkenylene" means unsaturatedhydrocarbon radicals of from 2 to 6 carbon atoms having one double bond,e.g. vinylene, propenylene, butenylene, pentenylene and hexenylenewherein the double bond may be present anywhere in the chain, e.g. 1- or2-propenylene, 1-, 2- or 3-butenylene. Similarly, "lower alkynylene"means a hydrocarbon radical of from 2 to 6 carbon atoms having onetriple bond, e.g. ethynylene, propynylene, butynylene, pentynylene andhexynylene, wherein the triple bond may be present anywhere in thechain, e.g. 1- or 2- propynylene, 1-, 2- or 3-butynylene.

Example of heterocyclic radicals are: ##STR15##

Compounds of the instant invention include various stereoisomers.Preferred stereoisomers are those in which the absolute configuration atcarbon atoms adjacent to both a nitrogen and a carbonyl groupcorresponds most closely to the absolute configuration of L-amino acids.

Preferred examples of compounds of formula I are as follows:

1-[2-(S)-[[1-(S)-carboxy-2-[4-[[[6-chloro-3-(chloromethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenylmethoxy}ethyl]amino]-1-oxopropyl]-[2S-(2α,3aα, 7aα)]-octahydro-1H-indole-2-carboxylic acid, S,S-dioxide,

1-[2-(S)-[[1-(S)-carboxy-2-[4-[[[6-chloro-3-(chloromethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenylmethoxy]ethyl]-(S)-alanyl-(S)-proline,S,S-dioxide,

1-[2-(S)-[[1-(S)-carboxy-3-[4-[[[6-chloro-3-(chloromethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]propyl]amino]-1-oxopropyl]-[2S-(2S-(2α,3aα, 7aα)]-octahydro-1H-indole-2-carboxylic acid, S,S-dioxide,

N-[1-(S)-carboxy-3-[4-[[[6-chloro-3-(chloromethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]propyl]-(S)-alanyl-(S)proline,S,S-dioxide,

N-[1-(S)-carboxy-4-[4-[[[6-chloro-3-(chloromethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7yl]sulfonylamino]methyl]phenyl]butyl]-(S)-alanyl-(S)proline,S,S-dioxide,

7-N-[2-(S)-[[1-(S)-carboxy-3-[4-[[[6-chloro-3-(chloromethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]propyl]amino]-1oxopropyl]-1,4-dithia-7-azaspiro,[4.4]nonane-8-(S)-carboxylicacid, S,S-dioxide,

N-[1-(S)-carboxy-5-[6-chloro-3-(chloromethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonyl-amino]pentyl-(S)-alanyl-(S)-proline,S,S-dioxide,

1-[2-(S)-[[1-(S)-carboxy-5-[[-6-chloro-3-(chloromethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]pentyl]amino]-1-oxopropyl]-[2S-(2.alpha.,3aα, 7aα)]-octahydro-1H-indole-2-carboxylic acid, S,S-dioxide,

and the 3-(dichloromethyl)-,3-(cyclopentylmethyl)-,3-(2-phenylethyl)-,3-(butyl)-,3-(pentyl)-, and3-(trifluoroethylthiomethyl)-1,2,4-benzothiazinyl analogues thereof.

The compounds of this invention form salts with various inorganic andorganic acids and bases which are also within the scope of theinvention. Such salts include ammonium salts, alkali metal salts, e.g.sodium and potassium salts, and alkaline earth metal salts, e.g. calciumand magnesium salts. Salts with organic bases may be prepared, e.g.,N-methylglucamine, lysine and arginine, as well as salts with organicand inorganic acids, e.g., HCl, HBr, H₂ SO₄, H₃ PO₄, methanesulfonicacid, toluenesulfonic acid, maleic acid, fumaric acid andcamphorsulfonic acid. The non-toxic pharmaceutically acceptable saltsare preferred, although other salts are also useful, e.g., in isolatingor purifying the product. The acid salts (e.g. HCl and maleate) arepreferred, especially the hydrochloride.

The salts may be formed by conventional means, as by reacting the freeacid or base forms of the product with one or more equivalents of theappropriate base or acid in a solvent or medium in which the salt isinsoluble, or in a solvent such as water which is then removed in vacuo,or by exchanging the cations of an existing salt for another cation on asuitable ion exchange resin.

Compounds of formula I may be prepared by several routes using methodsknown in the art.

For example, compounds of formula I can be prepared by condensing anacid of formula IV (or its hydrochloride salt) with an amino acidderivative of formula V: ##STR16## wherein R¹,R⁶, R⁷, R⁸, D, B, E and Aare as defined above. The reaction is carried out in an inert solventsuch as dimethylformamide (DMF) in the presence a condensing agent suchas 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide hydrochloride (DEC)and 1-hydroxybenzotriazole, and where the compound of formula IV is asalt in the presence of a base such as triethylamine. The reaction ispreferably carried out in an inert atmosphere at a temperature of 0°-25°C.

Compounds of formula V are known in the art, or may be prepared bymethods well known to those skilled in the art.

Compounds of formula IV may be prepared for example from the reaction ofa sulfonyl chloride of formula VI and an amine of formula VII: ##STR17##wherein D, B, E, R¹, R⁶ and R⁷ are as defined above and R¹⁴ is a readilyremovable ester protecting group such as t-butyl, benzyl ortrimethylsilylethyl. The reaction is carried out at 0°-5° C. in asolvent such as tetrahydrofuran (THF).

Compounds of formula VI may be prepared from know starting materialsusing procedures well known in the art. For example, when D is of theformula IIIa wherein G is SO₂ and R³ is phenylethyl, the sulfonylchlorides of formula VI may be obtained by reacting a disulfonylchloride of formula VIII with aqueous ammonia at low temperature(dry-ice-acetone bath) in a solvent such as 1,2-dimethyoxyetane (DME) inthe presence of a base such as triethylamine to obtain a sulfamide offormula IX, followed by reaction of the sulfoamide with phenyl propanalin a solvent such as DME and in the presence of an acid such asp-tolenesulfonic acid: ##STR18##

Similarly, when D is of formula IIIb, the sulfonyl chlorides of formulaVI may be obtained by well known procedures. A typical reaction schemewas follows: ##STR19##

Compounds of formula VII are prepared from known started materials usingmethods known in the art. A typical reaction scheme is shown below forcompounds of formula VIIb, wherein R¹ is hydrogen, R⁶ is lower alkyl, R⁷is methyl, J is --(CH₂)--, L is phenyl, M is --CH₂ --O--CH₂ --, E is--NH--, and R¹⁴ is as defined above: ##STR20##

Details of the above typical reaction scheme are disclosed in Example 1,Parts A-E.

In the above reaction scheme, the triflate reagent, i.e. t-butyl2(S)-trifluoromethanesulfonyloxy)propionate, reacts by nucleophilicdisplacement with the α-aminoacid ester to give a high yield of thecorresponding specific diastereomer of the resulting monoaminodicarboxylic acid ester.

Compounds of formula VIId, wherein M is (CH₂)₂ and R¹, R⁶, R⁷, R¹⁴, J, Land E are as described above for formula VIIb, may be prepared asfollows: ##STR21##

The above reaction scheme is exemplified in Parts A-C of Example 3.

A typical reaction scheme for the preparation of compounds of formulaVIIf wherein L is a bond, J is --(CH₂)_(s) -- and M is --(CH₂)_(u))--wherein the sum of s and u is 4, and R¹, R⁶, R⁷, R¹⁴ and E are asdescribed above for formula VIIb is as follows: ##STR22## Example 5provides details of this procedure.

Alternatively, intermediates of formulae VIIa or VIIc may be used toprepare compounds of formula I as follows: The R¹⁴ protecting groups(e.g. t-butoxycarbonyl) of compounds of formulae VIIa or VIIc may beremoved, e.g. by trifluoroacetic acid, and the nitrile reacted with anamino acid derivative of formula V under the conditions described onpage 12. The resulting nitrile can then be reduced to the correspondingamine, e.g., by hydrogenation, which amine can then in turn be coupledto a sulfonyl chloride of formula VI by conventional methods. Similarly,intermediates of formula VIIe (i.e., compounds of formula VII wherein Bis alkyl) may be deprotected at the carboxylic acid group and condensedwith the amino acid derivative of formula V as described above, thendeprotected at the amino group, e.g. by hydrogenation, and the resultantamine reacted with a sulfonyl chloride of formula VI by conventionalmethods.

Another method for preparing compounds of formula IV is that exemplifiedbelow for preparing compounds wherein D is of formula IIIa wherein G is--SO₂ --, J is --CH₂ --, is -aryl-Z- wherein aryl is phenyl and Z is##STR23## M is --CH₂ --, E is --NH--, R¹ is hydrogen, R⁶ is ethyl, R⁷ ismethyl, and R², R³ and R⁴ are as defined above: ##STR24##

Another method for the preparation of compounds of formula I involvesthe sulfonylation of an amine of formula X with a sulfonylchloride offormula VI: ##STR25## wherein R¹, R⁶, R⁷, R⁸, B, E and A are as definedabove. The reaction is carried out in an inert solvent such as THG inthe presence of a proton acceptor such as N-methylmorpholine. Thereaction is preferably carried out in an inert atmosphere at atemperature of 0°-25° C.

Compounds of formula X may be prepared by well known methods, forexample to obtain compounds wherein B is --CH₂ --C₆ H₅ --(CH₂)₂ --, E is--NH-- and R¹, R⁶, R⁷ and R⁸ are as defined above, compounds of formulaVII(c) may be deprotected at the terminal carboxy group and thencondensed with an amino acid of formula V: ##STR26## The nitrile offormula XII is reduced to an amine of formula X, e.g. by hydrogenation.

Compounds of formula I may also be prepared by condensing an aldehyde offormula XIII (or a reaction derivative thereof, e.g., an acetal) with anaminosulfonamide of formula XIV: ##STR27## wherein R¹, R³, R⁴, R⁶, R⁷,R⁸, B, E and A are as defined above. The reaction is carried out in aninert solvent such as THF in the presence of p-toluenesulfonic acid. Thereaction is preferably carried out in an inert atmosphere at atemperature of 0°-80° C.

Compounds of formula XIII are known in the art or may be prepared byknown methods.

Compounds of formula XIV can also be prepared by known methods, forexample a sulfonamide of formula IX can be reacted with an amine offormula VII to obtain a compound of formula XV: ##STR28## wherein R¹,R⁴, R⁶, R⁷, R¹⁴, B and E are as defined above. The protecting group R¹⁴is then removed e.g., by treating with hydrochloric acid in a solventsuch as dioxane. The resultant product is condensed with an amino acidof formula V under conditions similar to those described above for thereaction of compounds of formulae IV and V to obtain a compound offormula XIV.

Yet another process for the preparation of the compounds of the formulaI wherein X is sulfur comprises condensing a halide of the generalformula XV ##STR29## with a thiol of the general formula XVI ##STR30##where R¹, R⁶, R⁷, R⁸, A, E, J, L, t and v are as defined for formula Iand Hal represents halogen, preferably bromine. The reaction ispreferably carried out in an inert medium at a temperature of 0°-25° C.

Compounds of the formula XV may be prepared by well known methods.Illustrative of such methods, is the following specific reaction scheme:##STR31## Compounds of general formula XVI may also be prepared by knownmethods, the following specific reaction scheme being illustrative ofsuch methods: ##STR32##

The known coupling methods above include amino group protection duringthe coupling reaction, for example with protecting groups such asN-formyl, N-t-butoxycarbonyl (t-Boc) and N-carbobenzyloxy (Cbz) groups,followed by their removal to yield compounds of formula I. Furthermore,the COR⁸ function wherein R⁸ is OH may be protected by removable estergroups such as benzyl, ethyl, t-butyl, trimethylsilylethyl and the like.

The more complex esters at R⁶ (i.e., R⁶ is other than hydroxy or alkoxy)are most conveniently prepared by esterifying compounds of formula Iwherein R⁶ is hydroxy and R⁸ is a protected hydroxy, e.g. benzyloxy,with the appropriate reagents, e.g. chloromethyl pivalate in thepresence of base, to obtain the corresponding pivaloyloxymethyl ester;the benzyl group is then removed by conventional means, e.g. catalytichydrogenation.

When topically administered to the eye, the compounds employed in theinvention reduce intraocular pressure (IOP). For example,1-[2-(S)-[[1-(S)-carboxy-5-[[6-chloro-3-chloromethyl-3,4-dihydro-2H-1,2,4-benzothiadiazinyl-7-yl]sulfonylamino]-pentyl]amino]-1-oxopropyl]-[2S-(2.alpha.,3aα,7aα)]-octahydro-1H-indole-2-carboxylicacid, S,S-dioxide caused falls in IOP of a magnitude similar to thoseproduced by the anti-glaucoma agent timolol when each were administeredat concentrations of 0.00001 and 0.5 (w/v %), respectively, and testedby the following procedure:

Male New Zealand white rabbits having a normal IOP are conditioned tothe laboratory setting for at least one 4 hr period before being used tostudy drug effects. A Makay-Marg applanation tonometer is used tomeasure IOP. Readings, in mm Hg, are taken in triplicate and the averageis recorded.

Rabbits are restrained in a cloth sack 2 min. prior to IOPdetermination. The left lower eyelid is retracted to form a pouch and 1drop of a local anesthetic is irrigated over the cornea. The lowereyelid is then held closed over the eye for about 1 min. Cornealanesthesia is repeated before each set of IOP determinations. Readingsare taken just before dosing with drug (0 time) and at hourly intervalsthereafter. Drugs are administered in a 50 μl volume in the same manneras the anesthetic.

To prepare suitable dosage forms, the active compounds may beconveniently admixed with a non-toxic pharmaceutically acceptablecarrier suitable for topical ophthalmologic administration. Typical ofsuch pharmaceutically acceptable carriers are, for example, water,mixtures of water and water miscible solvents such as lower alkanols orvegetable oils, petroleum based jelly, and including also from 0.1 to 2%by weight of hyroxyethyl cellulose, ethyl oleate, carboxymethylcellulose, polyvinylpyrrolidone, and other water solubleophthalmologically acceptable non-toxic polymers, for example, cellulosederivatives such as methyl cellulose, alkali metal carboxymethylcellulose, hydroxyethyl cellulose, hydroxypropyl cellulose,hydroxypropylmethyl cellulose; acrylates such as polyacrylic acidssalts, ethylacrylates; polyacrylamides; natural products such asgelatin, alginates, pectins, tragacanth, karaya, chondrus, agar, acacia;the starch derivatives such as starch acetate, hydroxyethyl starchethers, hydroxypropyl starch, as well as other synthetic derivativessuch as polyvinyl alcohol, polyvinyl pyrrolidone, polyvinyl methylether, polyethylene oxide, neutralized carbopol and xanthan gum andmixtures of these polymers. The pharmaceutical preparation may alsocontain non-toxic auxiliary substances such as emulsifying, preserving,wetting, bodying agents and the like, as for example, polyethyleneglycols 200, 300, 400 and 600, carbowaxes 1,000, 1,500, 4,000, 6,000 and10,000, antibacterial components such as quaternary ammonium compounds;phenylmercuric salts known to have cold sterilizing properties and whichare non-injurious in use; thimerosal, methyl and propyl paraben, benzylalcohol, phenyl ethanol; buffering ingredients such as alkali metalchloride, borate, acetate, gluconate buffers; antioxidants such assodium metabisulfite, butylated hydroxyanisole (BHA), butylatedhydroxytoluene (BHT) and the like; and other conventional ingredientssuch as sorbitan monolaurate, triethanolamine, oleate, polyoxyethylenesorbitan monopalmitylate, dioctyl alkali metal sulfosuccinate,monothioglycerol, ethylenediamine tetracetic acid and the like.

Additionally, suitable ophthalmic vehicles can be used as carrier mediafor the present purpose including conventional phosphate buffer vehiclesystems, isotonic boric acid vehicles, isotonic alkali chloridevehicles, tris and the like.

The pharmaceutical preparation may also be in the form of a solidinsert. For example, one may use a solid water soluble polymer as thecarrier for the medicament. Inserts that are known in the art that aresuitable for this use include those described in British Patent 15611,and in U.S. Pat. Nos. 3,993,071; 3,986,510; 3,868,445; and 3,867,510.Solid water insoluble inserts, such as those prepared from ethylenevinyl acetate copolymer, may also be utilized.

The compositions of the invention may include therapeutically effectiveamounts of additional ophthamologically acceptable therapeutic agents inaddition to the compound of formula I. For example antibiotics andanesthetics, as well as other IOP lowering agents may be present.

The pharmaceutical compositions of the invention are administered in theform of ophthalmic pharmaceutical compositions adapted for topicaladministration to the eye; such as solutions, suspensions, ointments andsolid inserts. Formulations of the invention may contain from about0.000001% (w/v) to about 1.0% and especially about 0.00001% to about0.01% of a compound of formula I. As a unit dosage form, between about0.5 ng to about 0.05 mg preferably 5 ng to 5 μg of such compound isapplied to the human eye.

Where utilized herein, the term "controlling the elevated intraocularpressure" means the regulation, attenuation and modulation of increasedintraocular tension, which is the primary diagnostic symptom of thedisease glaucoma. The term also means that the diminution, in theotherwise elevated intraocular pressure, obtained by the practice of theinvention is maintained for a significant period of time as, forexample, between consecutive doses of the composition of the invention.

The compounds of formula I may be employed in the composition andmethods of the invention as the sole IOP lowering ingredient or may beused in combination (in the same or separate compositions) with othermechanistically distinct IOP lowering ingredients such asbeta-adrenergic blocking agents, (e.g., timolol). For purposes of thepresent invention, the term beta-adrenergic blocker means a compoundwhich by binding to beta adrenergic plasma membrane receptors reduces oreliminates sympathetic activity or blocks the effects of exogenouslyadministered catecholamines or adrenergic drugs. See, for example,Weiner, N., Drugs That Inhibit Adrenergic Nerves and Block AdrenergicReceptors, in The Pharmaceutical Basis of Therapeutics (ed. A. G.Goodman, L. S. Goodman, A. Gilman), Macmillan Publishing, New York,1980, 6th ed., pp. 188-197. Examples of preferred beta adrenergicblockers are bunolol(5-[3-(1,1-dimethylethyl)amino-2-hydroxypropoxy]-3,4-dihydro-1(2H)-naphthalenone),atenolol(4-[2-hydroxy-3-[(1methylethyl)amino]-propoxy]benzeneacetamide),metoprolol(1-[4-(2-methoxyethyl)phenoxy]-3-[(1-methylethyl)amino]-2-propanol),nadolol (5-[3-[(1,1-dimethylethyl)amino]-2-hydroxypropoxy]-1,2,3,4-tetrahydro-2,3-naphthalenediol), pindolol(1-(1H-indol-4-yloxy)-3-[(1-methylethyl)amino]-2-propanol), propranolol(1-[(1-methylethyl)amino]-3-(1-naphthalenyloxy)-2-propanol), timolol(1-[(1,1-dimethylethyl)amino]-3-[(4-morpholinyl-1,2,5-thiadiazol-3-yl)oxy]-2-propanol),labetalol(2-hydroxy-5-[1-hydroxy-2-[(1-methyl-3-phenylpropyl)amino]ethyl]-benzamide),betaxolol (1-[4-[2-(cyclopropylmethoxy)ethyl]-phenoxy]-3-[(methylethyl)amino]-2-propanol), carteolol(5-[3-[(1,1dimethylethyl)amino]-2-hydroxypropoxy]-3,4-dihydro-2(1H)-quinolinone),and dilevalol([R-(R,R)]-2-hydroxy-5-[1-hydroxy-2-[(1-methyl-3-phenylpropyl)amino]ethyl]-benzamide4-methylbenzenesulfonate salt), "and pharmaceutically acceptable saltsand isomers thereof".

The usefulness of beta adrenergic blockers for lowering intraocularpressure is known in the art. Thus, the beta adrenergic blocker timolol,is currently approved by the U.S. Food and Drug Administration fortopical use as a treatment for glaucoma. It is marketed in two dosestrengths, i.e., 0.25% and 0.5%. As previously stated, this agent mustbe used with caution in a defined patient population because ofrecognized untoward side effects (see Physicians Desk Reference forOphthalmology, 11th edition, 1983, p. 126, Medical Economics Co. Inc.,Oradell, N.J. 07649).

As one aspect of the present invention, it is contemplated that areduction in intraocular pressure equivalent to that obtained by use ofa beta-blocker, e.g., the approved clinical dose of the beta-blockertimolol, may be obtained by use of a lower dose of beta-blocker whensuch lower dose is combined with an effective amount of a compound offormula I. It is anticipated that the use of the diminished dosage ofbeta-blocker, e.g., timolol, will result in a reduction of severity andfrequency of timolol-like related side effects.

For purposes of this combination treatment modality, the beta-blockerand compound of formula I are preferably administered simultaneously asone composition in one pharmaceutical dosage form, but they may beapplied as separate topical compositions, if desired. When applied aspart of a composition including a compound of formula I, the betaadrenergic blocker may comprise from about 0.01% to about 1.0% of thecomposition of the invention. The preferred ranges of the components inthe composition of the invention in unit dosage form are as follows:

Beta adrenergic blocker: from 5 μg to 250 μg

Compound of formula I: from 5 ng to 5 μg.

When applied in separate compositions, the beta-adrenergic blocker andcompound of formula I may be included in such compositions in the sameranges. The individual dosage requirements, i.e., the amount of eachdose and the frequency of administration, may vary depending on theseverity of the disease and the response of the patient.

Since the composition of the invention and the composition including thebeta-adrenergic blocker can be administered separately, the inventionalso relates to combining separate pharmaceutical compositions in kitform, that is, combining two separate units, an inventive pharmaceuticalcomposition and a topical pharmaceutical composition including abeta-adrenergic blocker, in a single package. A particular advantage ofthe kit resides in the ability to provide a combination of an inventivecomposition which can be administered once or twice a day and a topicalbeta-adrenergic blocker composition which may be administered asnecessary or desired.

Those skilled in the art will appreciate that the "intraocular pressurereducing concentration" for such combination therapy will consist of arange of concentrations (doses), and that there will be a lower limit tosaid concentration below which, the present invention will not operate.For purposes of this invention, this lower limit or minimum dosage maybe considered to be about 5% of the effective dose (threshold dose) ofthe particular component. The intraocular pressure reducingconcentration that is actually utilized, whether for a compound definedin formula I or for a particular beta adrenergic blocker, will dependon, inter alia, the potency of each particular material, the combinationbeing administered and the age, size and condition of the patient beingtreated as well as on the severity of the disease state.

We also contemplate that the elevation in IOP associated with theclinical ophthalmic and systemic use of anti-inflammatory steroids canbe reduced by the administration of a composition of the presentinvention. In particular, an increase in IOP is most often associatedwith the administration of steroidal anti-inflammatory agents.Anti-inflammatory steroids include hydrocortisone, cortisone,prednisolone, orednisone, dexamethasone, methylprednisolone,triamcinolone, betamethasone, alclometasone, flunisolide,beclomethasone, clorocortolone, diflorasone, halcinonide, fluocinonide,flucinolone, desoximetasone, medrysone, paramethasone,9,21-dichloro-17-[(2-furanylcarbonyl)oxy]-11-hydroxy-16α-methyl-pregna-1,4-diene-3,20-dioneand fluorometholone, and their pharmaceutically acceptable salts andesters. Preferred steroids are hydrocortisone, prednisolone,dexamethasone, betamethasone, beclomethasone, medrysone andfluoromethalone and their pharmaceutically acceptable salts and esters.This rise in IOP may occur with all modes of administration of thedrugs, including systemic (usually oral), local injection (e.g., depotinjection), and especially with ophthalmic topical or intravitrealadministration. The composition of the present invention may beadministered following steroid treatment to lower elevated IOP, or maybe co-administered with the steroid to suppress the IOP-raising effectof the steroid while not interfering with the anti-inflammatory activityof the steroid.

It is further contemplated that any possible combination of dosage formsmay be used to administer the combination, e.g., oral steroid/topicalcomposition of the invention, topical steroid/oral composition of theinvention, oral steroid/oral composition of the invention, topicalsteroid/topical composition of the invention, and locally injectedsteroid/topical composition of the invention, although a preferredcombination comprises a steroid and a topical composition of theinvention. For ophthalmic use, a combination of a topical steroid and atopical composition of the invention is preferred. More preferred is atopical ophthalmic pharmaceutical dosage form comprising both a steroidand a composition of the invention. Such compositions or combinationscan be employed in a method for reducing and controlling the elevatedIOP associated with-ophthalmic and systemic use of steroidalanti-inflammatory agents, which method comprises administering to amammal effective amounts of a steroid and a composition of theinvention, either separately or in the same pharmaceutical composition.

Since the present invention relates to treatment with a combination of acomposition of the invention and a steroidal anti-inflammatory agentwherein the inventive composition and steroid may be administeredseparately, the invention also relates to combining separatepharmaceutical compositions in kit form, that is, combining two separateunits, an inventive pharmaceutical composition and a steroidpharmaceutical composition, in a single package. Preferred components ofthe kit comprise a topical opthamological pharmaceutical compositionincluding a compound of formula I and a pharmaceutically acceptablesteroid composition. More preferred components of the kit are a topicalopthamological inventive pharmaceutical composition including a compoundof formula I and a topical opthamological steroid pharmaceuticalcomposition. A particular advantage of the more preferred embodiment ofthe kit resides in the ability to provide a combination of an inventivecomposition which can be administered once or twice a day and a steroidcomposition which may be administered as frequently as once each hour.

In this combination treatment modality, topical formulations of theinvention may combine the following amounts of each inventivecomposition and steroidal constituent, or each constituent may beadministered separately:

A compound of formula I from about 0.000001% (w/v) to about 1.0% andespecially about 0.00001% to about 0.01% of medicament. As a unit dosageform, an amount of a compound of formula I from about 0.5 ng to about0.5 mg preferably about 5 ng to about 5 μg, of the active component isapplied to the human eye. Individual dosage requirements, i.e., theamount of each dose and the frequency of administration, will depend onthe potency of the particular composition of the invention, the severityof the increase in IOP and the response of the patient.

Steroid from about 0.05 to about 1.5 (w/v %) of medicament. As a unitdosage form, an amount of steroid from between 20 μg to 600 μg of theactive composition is applied to the human eye. Individual dosagerequirements, i.e., the amount of each dose and the frequency ofadministration will depend on the potency of the particular steroid, theseverity of the disease and the response of the patient. Approximateranges for such steroids are well known to those skilled in the art. Theparticular steroid selected will determine which inventive compositionand concentration thereof to select for use in a combinationpreparation.

In one embodiment of the invention, both active ingredients, i.e.,inventive composition and steroid, will be administered simultaneouslyand be contained in one pharmaceutical dosage form, each component beingpresent in the dosage form in its own respective preferredconcentration. When the steroid is administered systemically ortopically other than in an ophthalmological composition, theconcentration of the steroid in the composition and the unit dosageweight may vary considerably, depending as above on such factors as thepotency of the steroid, its onset and duration of action as well as theseverity of the disease, and the response of the patient. Appropriatedosage ranges for systemic and topical administration of each steroidare well known in the art.

Those skilled in the art will know that for solutions and suspensions, aparticular dosage of active ingredient may be calculated if one assumesthat one drop of solution is being administered and if one knows theconcentration (w/v) of the particular solution that is beingadministered. Thus, one drop (1/20 ml) of a 0.25% solution (contains 2.5mg of active per ml) is known to contain .125 mg or 125 μg of activeingredient.

The IOP-lowering effects of the compositions employed in the inventionmay be measured by the procedure described by Watkins et al., J. OcularPharmacol. 1 (2):161-168, 1985.

The following procedures and examples illustrate the preparation ofcompounds employed in this invention.

PREPARATION 1 t-BUTYL 2(S)-(TRIFLUOROMETHANESULFONYLOXY)PROPRIONATE(Triflate Reagent)

A. Add 2(S)-(p-toluenesulfonyloxy)propionic acid (4.4 g) to a coldsolution of 10ml of isobutylene and 0.4 ml of concentrated sulfuric acidin 30 ml of methylene chloride in a pressure vessel, seal, and agitateat room temperature for 48 hours. Pour into 50 ml of 15% sodiumcarbonate solution, dry over magnesium sulfate and concentrate to obtaint-butyl 2(S)-(p-toluenesulfonyloxy)propionate as an oil (NMR δ 1.37).Distilled material (Kugelrohr, 120°) has [α]D²⁶ =-45.9° (EtOH, c=1).

B. Combine the product of part A (100 g) with acetic acid (40.0 g) andtriethylamine (67.2 g) in 200 ml of dry DMF. Heat at 65° for 20 hours.Partition with 21 each ether and water, and wash the ether with citricacid, then with dilute sodium bicarbonate solution. Dry and concentratethe ether solution to obtain t-butyl 2(R)-acetoxypropionate as acolorless liquid, bp 50° C./0.1 mm.

C. Combine the product of part B (62,6 g) with ethylenediamine (11.6 g)and heat at 70° for 24 hours. allow to cool, add 300 ml ether andfilter. Wash the ether with water, 10% citric acid, and then with sodiumbicarbonate solution. Dry and concentrate the ether solution to leave acolorless oil. Crystallize from hexane at -20° to give t-butyl2(R])-hydroxypropionate as white needles, m.p. 41°-2° C.

D. Combine the product of part C (10 g) with pyridine (6 ml) in 100mlmethylene chloride. Cool to -70° C., and add dropwise over a period ofabout 45 minutes a solution of trifluoromethanesulfonic anhydride (13.5g) in 20 ml methylene chloride, maintaining the temperature below -15°C. Stir at -20° C. for 30 min. Add ether and wash successively withwater, 4 of aq HCl, sat'd NaHCO₃ and brine. Dry and concentrate theorganic layer to obtain the title compound.

PREPARATION 26-CHLORO-3,4-DIHYDRO-3-(2-PHENYLETHYL)-2H-1,2,4-BENZOTHIADIAZINE-7-SULFONYLCHLORIDE 1,1-DIOXIDE

A. Dissolve 5 g 2-chloroaniline-3,5-disulphonyl chloride in 20 ml DME,cool to in a dry-ice/acetone both and add 2 ml triethylamine.

Add dropwise 25% ammonium hydroxide in water (1 ml) in DME (4 ml), stirat in a dry-ice acetone bath for 1 hour, allow to warm to roomtemperature, and stir for 90 min. Dilute the resultant reaction mixturewith ethyl acetate, wash with 4% ac HCl, water and brine, dry over MgSO₄and evaporate to obtain a solid residue.

B. Combine 13.6 g of the sulfonamide prepared in Step A, 6.57 g phenylpropanal, 25 ml DME and 20 mg p-tolulenesulfonic acid and stir at roomtemperature under N₂ for 3 hours. Evaporate the solvent, dissolve theresultant residue in 250 ml ethyl acetate, wash with 100 ml sat'd aq.NaHCO₃, and 100 ml brine, then dry over MgSO₄, filter and evaporate thesolvent to obtain the crude title compound. Purify the crude residue byprecipitation in CH₂ Cl₂ ; mp. 167.0°-167.5° C.

PREPARATION 3 CIS,SYN-OCTAHYDROINDOLE-2(S)-CARBOXYLIC ACID, t-BUTYLESTER

A. Dissolve the product of Preparation 4 (77 g) in absolute ethanol (900ml), add 5% Pd/C (10 g) and hydrogenate at room temperature at aninitial pressure of 60 p.s.i. After 3 hours, filter off the catalyst andwash with hot methanol. Evaporate the combined filtrate and wash invacuo, triturate the resultant residue in ethanol (100 ml), chill thesolution, then filter and air dry the resultant precipitate to obtain aresidue, m.p. 269°-270° C.

B. Suspend the product of Part A in dioxane (400 ml) and conc. H₂ SO₄(40 ml), add isobutylene (300 ml) and shake in a Parr shaker for 28hours. Pour the resultant reaction mixture into 50% aqueous NaOH (150ml) in 500 ml ice water and extract with ethyl ether (3×500 ml). Washthe combined organic extracts with water, then brine. Dry the organiclayer over Na₂ SO₄ and evaporate the solvent to obtain the titlecompound.

PREPARATION 4 CIS,SYN-OCTAHYDROINDOLE-2(S)-CARBOXYLIC ACID BENZYL ESTER

A. Dissolve 27.0 g of ethyl indole-2-carboxylate in 250 ml oftrifluoroacetic acid. Add 2.05 g of platinum oxide, hydrogenate themixture at 50 lb/in² at room temperature. Filter the mixture andconcentrate the filtrate in vacuo to give a residue. Suspend the residuein ether and treat with cold dilute sodium hydroxide solution. Dry theorganic layer over magnesium sulfate and concentrate it to give ethyloctahydroindole-2-carboxylate, a pale yellow oil.

B. Dissolve 116 g 10-d-camphorsulfonic acid in 1 liter of warm ethylacetate and add a solution of 86 g of the product of part A in 1 literof ethyl acetate. Allow the mixture to crystalline, heat to reflux, coolto room temperature, and filter. Recrystallize the filter cake from amixture of 500 ml isopropanol and 1800 ml ethyl acetate, filter and drythe crystals to obtain 2-(S)-carboethoxy-cys,syn-octahydroindole,d-10-camphorsulfonate, m.p. 192°-193° C.

C. Heat the product of Part B (107.6 g) and d-10-camphor-sulfonic acid(6.35 g) in benzyl alcohol (270 ml) at 105° C. under vacuum for 6 hoursor until TLC (silica, elute neutralize sample with ethyl ether)indicates reaction is complete. Pour the resultant residue into ethylether, seed and stir to obtain a precipitate. Filter the precipitate,wash with ethyl ether (2×500 ml) and dry the resultant residue undervacuum to obtain 2-(S)-benzyloxy-cis, syn-octahydro-indole,d-10-camphorsulfonate, m.p. 114°-118° C.

D. Suspend the product of Part C (150 g) in ethyl ether (1500 ml), add 1N aqueous NaOH (300 ml) and stir until the solid dissolves. Separate theorganic layer and wash the aqueous layer with ethyl ether (2×200 ml).Combine the organic layer, wash with brine, dry over Na₂ SO₄ andevaporate the solvent to obtain the title compound.

PREPARATION 56-CHLORO-3,4-DIHYDRO-3--(CHLOROMETHYL)-2H-1,2,4-BENZOTHIADIAZINE-7-SULFONYLCHLORIDE 1,1-DIOXIDE

A. Dissolve the sulfonamide prepared in Part A of Preparation 2 (20 g)in dry DME (100 ml), add chloroacetaldehyde dimethyl acetal (10 ml) andp-toluenesulfonic acid and reflux for 3 hours or until TLC (silica, 3%ethyl acetate in CH₂ Cl₂) indicates reaction is complete. Evaporate thesolvent, dissolve the resultant residue in ethyl acetate, wash withsaturated NaHCO₃, then brine and concentrate to half volume. Refrigerateovernight, filter the resultant precipitate, wash in hexane, filter anddry to obtain the title compound.

EXAMPLE 11-[2-(S)-[[1-(S)-CARBOETHOXY-2-[4-[[[6-CHLORO-3,4-DIHYDRO-3-(2-PHENYLETHYL)-2H-1,2,4-BENZOTHIADIAZIN-7YL]SULFONYLAMINO]METHYL]PHENYLMETHOXY]ETHYL]AMINO]-1OXOPROPYL]-[2S-[2α, 3aα, 7aα)]-OCTAHYDRO-1H-INDOLE-2-CARBOXYLIC ACID,S,S-DIOXIDE

A. To 10.4 g NaH (50% in mineral oil, washed with hexane) in 50 ml DMFat 0°-5° C., add dropwise over a 1 hour period 20o ofN-a-t-butoxycarbonyl-L-serine in 350 ml DMF. Stir at room temperaturefor 1 hour, then at 45° for 1 hour. Cool the reaction mixture to 0°-5°C. and add dropwise over 30 minutes 21.3 g of p-cyanobenzylbromide in100 ml DMF. Stir at 0° C. for 80 minutes, add 30 ml water, stir andfilter. Concentrate the filtrate and partition between ethyl acetate andsat'd. aq. NaHCO₃ /H₂ O. Wash the aqueous phase with ethyl acetate,adjust to pH 7.5 with 6 N HCl and concentrate to approximately 100 mls.

B. To the product of Step A, add 60 ml methanol, 40 ml ethyliodide, and4 g NaHCO₃. Stir under a nitrogen atmosphere for 72 hours, evaporate thesolvent in vacuo and partition the residue between 800 ml ethyl acetateand 800 ml water. Separate the organic layer, and extract the aqueouslayer with ethyl acetate; combine the organic extracts, wash with brine,dry over MgSO₄, filter and evaporate the solvent. Purify the resultantresidue by High Pressure Liquid Chromatography (HPLC) using 2 Prep 500cartridges and eluting with 21% ethyl acetate in hexane. Combine thedesired fractions and evaporate the solvent to obtain a residue. FABmass spec m/e=349 (M+H).

C. Cool 2 g of the product of Step B to 0°-5° C. and add dropwise 25 mlof trifluoroacetic acid. Let stand until TLC (silica, elute withhexane:ethyl acetate) indicates no starting material is left. Add ethylacetate, then evaporate the solvent in vacuo. Dissolve the resultantresidue in ethyl ether, and wash with 1 N aqueous NaOH; backwash theaqueous phase with ether, combine the ethereal extracts, dry over K₂ CO₃and evaporate the solvent to obtain a residue.

D. Cool 1.6 g triflate reagent (Preparation 1) in 10 ml CH₂ Cl₂ to 0° C.Add dropwise 1.1 g of the product of Step C and 1.2 g of PROTON SPONGE®(1,8-bis(dimethylamino)naphthalene, Aldrich Chemical Co., Milwaukee,Wis.) in 20 ml CH₂ Cl₂. Monitor reaction by TLC, adding1,8-bis-(dimethylamino)naphthalene and triflate reagent as necessary.Filter the resultant precipitate with the aid of ethyl acetate,evaporate the solvent, and purify the resultant residue by columnchromatography, eluting with 30% ethyl acetate in hexane. FAB mass specm/e=377 (M+H).

E. Dissolve 2.5 g of the product of Step D in 50 ml ethanol saturatedwith NH₃, add 1.25 g 5% Rh/Al₂ O₃ and hydrogenate at 60 psi for 4 hours.Filter the resultant mixture through celite with the aid of ethanol,then evaporate the solvent in vacuo to obtain a residue.

F. Cool to 0° C. 2.5 g of the product of Step E in 25 ml dry THF and adddropwise 3.3 g6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazine-7-sulfonylchloride 1,1-dioxide in 20 ml dry THF. Stir at 0° C. for 1 hour, thenadd 0.6 ml N,N-diisopropylethylamine and stir at room temperature for 2hours. (Reaction may be monitored by TLC [silica; elute with hexane:ethyl acetate]). Add the reaction mixture to ethyl acetate, wash with 4%aq. HCl, sat'd NaHCO₃ and brine, then dry over MgSO₄ and evaporate thesolvent in vacuo. Purify the resultant residue by HPLC: dissolve theresidue in acetone:ethyl acetate:hexane (20:35:45) and separate on 2Prep 500 cartridges using acetone:ethyl acetate:hexane (5.5:36.5:58) asmobile phase. Monitor eluent by TLC (silica; elute with acetone:ethylacetate:hexane [6:39:55]), combine the desired fractions and evaporatethe solvent in vacuo to obtain a residue. FAB mass spec m/e=766 (M+H).

G. Stir 2.9 g of the t-butyl ester of Step F in 40 ml HCl/dioxaneovernight; pass nitrogen through the solution to evaporate the solventand obtain the free acid.

H. Dissolve 2.9 g of the produce of Step G in 6 ml DMF, add 1.1 gcis,syn-octahydro-1H-indole-2(S)carboxylic acid, t-butyl ester and 700mg 1-hydroxybenzotriazole. Cool to 0° C., add 0.7 ml triethylamine and900 mg 1-(3-dimethylaminopropyl)-3-ethyl carboiimide hydrochloride andstir overnight. Evaporate the solvent in vacuo, take up the residue inethyl acetate, and wash with water, 4% aq. HCl, sat'd aq. NaHCO₃, andbrine. Dry the organic layer over MgSO₄, filter and evaporate to obtaina residue.

Purify said residue by HPLC using 2 Prep 500 cartridges andmethanol:ethyl acetate:hexane (5.25:36:58.75) as mobile phase (residuedissolved in methanol:ethyl acetate:hexane [10:35:55]). Combine thedesired fractions as determined by TLC and evaporate the solvent.Rechromatograph the resultant residue by HPLC usingaceone:ethylacetate:hexane (10:40:50) as mobile phase, combining thedesired fractions and evaporating the solvent to yield the titlecompound as a t-butyl ester. FAB mass spec m/e=917 (M+H).

I. Stir 1.5 g of the product of Step H in 20 ml dioxane saturated withHCl overnight; pass nitrogen through the solution to evaporate thesolvent. Purify the resultant residue on Dowex Ag 50 2X (H⁺ form),eluting with (ethanol:water [1:1]):pyridine (95:5). Combine desiredfractions as determined by TLC (silica; CH₂ Cl₂ :MeoH:AcOH [90:5:3]) andevaporate the solvent to obtain the title compound. FAB mass specm/e=861 (M+H).

EXAMPLE 21-[2-(S)-[[1-(S)-CARBOXY-2-[4-[[[6-CHLORO-3,4-DIHYDRO-3-(2-PHENYLETHYL)-2H-1,2,4-BENZOTHIADIAZIN-7-YL]SULFONYLAMINO]METHYL]PHEYLMETHOXY]ETHYL]AMINO]-1OXOPROPYL]-[2S-(2α, 3aα, 7aα)]-OCTAHYDRO-1H-INDOLE-2-CARBOXYLIC ACID,S,S-DIXOIDE.

A. Treat the produce of Example 1, Step G in a manner similar to thatdescribed in Example 1, Step H, first paragraph, substitutingcis,syn-octahydro-1H-indole-2(S)-carboxylic acid, benzyl estercamphorsulfonate salt (see Preparation 4, part C) for the t-butyl ester.

Purify the resultant residue by column chromatography on 100 g silicaeluted with CHCl₃ :ethyl acetate. Combine the desired fractions asdetermined by TLC (silica; elute with CH₂ Cl₂ :methanol:acetone[93:2:5]) and evaporate the solvent to obtain a residue. Further purifythe residue on a sephadex column (350 g). FAB mass spec m/e=951 (M+H).

B. Suspend 450 mg of the diester obtained in Step A in 2 ml water. Add 2ml 1 N aq. NaOH and stir overnight at room temperature. Adjust to pH 6-7with 1 N HCl, filter the resultant solid and dry under vacuum to obtainthe title compound. FAB mass spec m/e=833 (M+H).

EXAMPLE 31-[2-(S)-[[3-[4-[[[6-CHLORO-3,4-DIHYDRO-3-(2-PHENYLETHYL)-2H-1,2,4-BENZOTHIADIAZIN-7-YL]SULFONYLAMINOMETHYL]PHENYL]-1-(S)-(ETHOXYCARBONYL)PROPYL]AMINO]-1OXOPROPYL]-[2S-(2α,3aα, 7aα]-OCTAHYDRO-1H-INDOLE-2-CARBOXYLIC ACID, S,S -DIOXIDEHYDROCHLORIDE A. 2-Amino-4-(4-cyano)phenylbutanoic acid, ethyl ester

Reflux 2-(4-cyano)phenylethyl bromide (23 gm), the p-chlorobenzaldimineof ethyl glycinate (21 gm), tetra-n-butylammonium bromide (10 oms) andfreshly ground fine potassium carbonate powder (42 gms) in acetonitrile(150 mls) with mechanical stirring under nitrogen for 12 hours.

Cool the mixture, filter off the solid and wash the filter cake withethyl acetate (3×150 mls). Wash the combined filtrate with water (2×100mls) and evaporate the solvent in vacuo. Stir the residue vigorouslywith THF (200 mls) and 2 N HCl (200 mls) at room temperature for 2hours. Wash the aqueous phase with ethyl acetate, basify with solidpotassium carbonate to pH 9 and extract with ethyl acetate to give thetitle compound of Part A.

B. N-[1-(S)-(Ethoxycarbonyl)-3-(4-cyano)phenyl]-(S)-alanine, t-butylester

Slowly add the product of Part A (9 gms) and PROTON SPONGE® (17.2 gm) indry dichloromethane (80 mls) dropwise into a stirred solution oftriflate reagent (22 gm) in dry dichloromethane (40 mls) cooled in anacetone-ice bath. Stir at room temperature overnight. Filter theresultant precipitate and wash the filter cake with ethyl acetate (5×100ml). Wash the combined filtrate with 10% citric acid (3×100 ml); sodiumbicarbonate (sat'd, 2×100 ml), and saturated brine (2×100 ml). Dry thesolution over potassium carbonate in the presence of triethyl amine (5mls), and remove the solvent in vacuo. Chromatograph the resultantresidue (hexane:EtOAc:CH₂ Cl₂ [8:1:1], 1% Et₃ N; 500 gm silica gel;230-400 mesh) to obtain the title compound of Part B.

C. N-[3-(4-Aminomethyl)phenyl-1-(S)-(ethoxycarbonyl)propyl]-(S)-alanine, t-butyl ester

Hydrogenate a mixture of the product of Part B (2 g), hydrogen chloride(0.2 gm) and 10% Pd/C (0.4 gm) in absolute ethanol (100 ml) at 50 psifor 5 hours. Filter the resultant mixture through celite. Evaporate thesolvent to obtain the hydrogen chloride salt of the title compound ofPart C.

D. Add N-methylmorpholine (0.5 ml) to a solution of the product of PartC (1 gm) in dry THF (20 mls) cooled in acetone-ice bath (-5° C). Add thesulfonyl chloride of Preparation 2 (1.3 gm) and stir the resultingmixture at room temperature overnight. Dilute the resultant reactionmixture with ethyl acetate (400 ml), wash with 0.5 N HCl (100 ml),saturated NaHHCO₃ (2×100 ml), and brine (2×100 ml). Dry the solutionover MgSO₄ and remove the solvent in vacuo. Purify the resultant residueby chromatography [400 gm silica gel, 230-400 mesh; firsthexane:EtOAc:CH₂ Cl₂, 4:1:1, then hexane:EtOAc:CH₂ Cl₂, 1:1:1] to obtaina residue.

E. Add the product of Part D (1.76 gm) to 5.5 M HCl in dioxane (50 ml)and stir the resulting mixture at room temperature overnight. Evaporatethe solvent in vacuo, triturate the solid residue with ether, and removethe solvent in vacuo to obtain a residue (hydrogen chloride salt).

F. Treat the product of part E in a manner similar to that described inExample 1, Part H, first paragraph, substituting N-methylmorpholine fortriethylamine to obtain a residue. Purify the resultant residue bychromatography [400 gm silica gel 230-400 mesh; hexane:EtOAc:CH₂ Cl₂,1:2:1] to obtain the t-butyl ester of the title compound.

G. Stir the product of Part F (0.9 gm) in 5.5 M HCl/dioxane (40 mls) atroom temperature for 2 hours. Remove the solvent in vacuo, triturate theproduct with ether, and dry in vacuo to obtain the title compound ofExample 3.

EXAMPLE 41-[2-(S)-[[1-(S)-CARBOXY-3-[4-[[[6-CHLORO-3,4-DIHYDRO-3-(2-PHENYLETHYL)-2H-1,2,4-BENZOTHIADIAZIN-7-YL]SULFONYLAMINO]METHYL]PHENYL]-PROPYL]AMINO]-1-OXOPROPYL]-[2S-(2α,3aα,7aα)]-OCTAHYDRO-1H-INDOLE-2-CARBOXYLICACID, S,S- DIOXIDE

Dissolve the product of Example 3 (0.85 gm) in methanol (2 mls) and coolto 0° C under nitrogen. Add 1 N sodium hydroxide (5 mls) portionwise.Refrigerate the mixture overnight. Acidify the resultant mixture withacetic acid, evaporate to dryness, and purify the residue bychromatography on a C-18 medium pressure reverse-phase column to yieldthe title compound of Example 4.

EXAMPLE 51-[[2-(S)-[5-[[6-CHLORO-3-CHLOROMETHYL-3,4-DIHYDRO-2H-1,2,4-BENZOTHIADIAZIN-7-YL]SULFONYLAMINO]-1-(S)-(ETHOXYCARBONYL)PENTYL]AMINO]-1-OXOPROPYL]-[2S-(2α,3aα,7aα)]-OCTAHYDRO-1H-INDOLE-2-CARBOXYLICACID, S,S-DIOXIDE

A. Combine N.sup.ε Cbz lysine, ethyl ester (30.0 g), t-butylbromopropionate (44.78 g), triethylamine (14.8 ml) and DMF (120 ml) andstir under nitrogen at 75° C until TLC shows no starting materialpresent. Evaporate the solvent, dilute the resultant residue with waterand extract with ethyl ether. Wash the organic layer with brine, dryover MgSO₄, filter and evaporate the solvent in vacuo.

Purify the resultant residue by column chromatography on 1,100 g silicagel (60-200 mesh), eluting with ether:hexane (50:50.75:50). Combine thedesired fractions and evaporate the solvent in air. Further purify theresultant residue by HPLC using 2 Prep 500 cartridges and eluting withether:hexane. Combine desired fractions, dry over MgSO₄ and evaporatethe solvent in air to obtain a residue.

B. Combine 20% Pd(OH)₂ /C (21.1 g) and anhydrous ethanol (25 ml) in aParr shaker bottle, add the product of Part A (16.5 g) in ethanol (55ml) and hydrogenate overnight under 60 p.s.i. H₂. Filter the resultantsolution over filter paper and celite and evaporate the solvent toobtain a residue.

C. Dissolve the sulfonyl chloride prepared in Preparation 5 (2.4 g) indry DME (15 ml). Add triethylamine (1 ml) and the product of Part B ofthis Example (2 g) in DMF (10 ml) and stir for 1 hour, or until TLC(silica, 10% MeOH in CH₂ Cl₂) indicates no starting material is left.Add the resultant solution to ethyl acetate, wash with water, saturatedNaHHCO₃ and brine, dry over MgSO₄ and evaporate the solvent in vacuo.Purify the resultant residue on a Sephadex LH20 column to obtain aresidue. FAB mass spec m/e=632 (M+H).

D. Treat the product of Part C in a manner similar to that described inExample 1, Part G.

E. Treat the product of Part D in a manner similar to that described inExample 1, Part H, first paragraph, substituting the camphorsulfonatesalt of the benzyl ester of the octahydroindole (see Preparation 4 PartC) for the t-butyl ester.

Purify the resultant residue on a sephadex LH20 column, combine thedesired fractions and evaporate the solvent. Dissolve the resultantresidue in ethyl acetate, add dioxane saturated with HCl and evaporatethe solvent to obtain the benzyl ester of the title compound. FAB massspec m/e=817 (M+H).

F. Dissolve the product of Part E (600 mg) in acetic acid saturated withhydrogen bromide (6 ml). After 5 hours, evaporate the solvent and purifythe resultant residue on a Sephadex LH20 column. Combine the desiredfractions as determined by TLC (silica, MeOH:CH₂ Cl₂ :acetic acid,10:90:4) and evaporate the solvent to obtain the title compound. FABmass spec m/e=726 (M+H).

EXAMPLE 61-[[2-(S)-[1-(S)-CARBOXY-5-[[6-CHLORO-3-CHLOROMETHYL-3,4-DIHYDRO-2H-1,2,4-BENZOTHIADIAZIN-7-YL]SULFONYLAMINO]PENTYL]AMINO]-1-OXOPROPYL]-[2S-(2α,3aα,7aα)]-OCTAHYDRO-1H-INDOLE-2-CARBOXYLICACID, S,S-DIOXIDE

Add the product of Example 5, Part E (benzyl ester) to 1 N aqueous NaOH(4 ml) and water (4 ml) and stir overnight. Add 1 N HCl (4 ml) andethanol. Charge the resultant solution to Dowex Ag 50 cation exchangeresin by stirring batchwise for 20 minutes (60 ml resin, pre-washed withethanol:water, 1:4). Prepare a column from the loaded resin, elute thecolumn with ethanol:water (1:4) until the elute is pH 6, then elute withethanol/water:pyridine (95:5). Combine the desired fractions asdetermined by TLC (silica, ethanol:water, 9:1). Further purify theresultant product on a Sephadex LH 20 column. Combine the desiredfractions and evaporate the solvent to obtain the title compound. FABmass spec m/e+698 (M+H).

EXAMPLE 7N-[2-[[4-[[[6-CHLORO-3,4-DIHYDRO-3-(2-PHENYLETHYL)-2H-1,2,4-BENZOTHIADIAZIN-7-YL]SULFONYLAMINO]METHYL]PHENYL]CARBONYL]AMINO-1-(S)-(ETHOXYCARBONYL)ETHYL]-(S)-ALANYL-(S)-PROLINE,S,S-DIOXIDE

A. Dissolve 4-cyanobenzoic acid, t-butyl ester (11.34 g) in ethanol (100ml) saturated with anhydrous NH₃, add 5% Rh/Al₂ O₃ (120 g) andhydrogenate in a Parr apparatus at 60 p.s.i. at room temperature for23/4 hours. Filter the resultant solution through celite and evaporatethe solvent to obtain a residue.

B. Dissolve the product of Part A (11.27 g) in dry THF (100 ml) addN,N,-diisopropylethyl amine (8.44 g) and cool to 0° C. in an ice bath.Add dropwise, slowly and with stirring, the sulfonyl chloride preparedin Preparation 2 (27.51 g) and let stand at 0° C. for 35 minutes. Removethe ice bath and stir at room temperature for 2 1/2 hours or until TLC(silica, CH₂ Cl₂ :MeOH, 95:5) shows the reaction to be completeEvaporate the solvent to obtain a residue.

C. Cool to 0° C. a solution of the product of Part B (3.00 g) in CH₂ Cl₂(25 ml) and add, slowly and with stirring, trifluoroacetic acid (25 ml).Stir for 30 minutes at 0° C., then at room temperature for 21/2 hours oruntil TLC (as in Part B) shows the reaction to be complete. Evaporatethe solvent to obtain a residue.

D. Dissolve triflate reagent as prepared in Preparation 1 (3.31 g) inCH₂ Cl₂ (75 ml) and cool to 0° C.; add, slowly and with stirring asolution of PROTON SPONGE® (4.24 g) and N-βCbz-2,3-diaminopropionate,ethyl ester (2.00 g) in CH₂ Cl₂ (75 ml). Stir at 0° C. for 15 hours.Extract the solution with 10% citric acid (2×), then sat'd. NaHHCO₃(2×), dry the organic layer over MgSO₄, filter and evaporate thesolvent. Purify the resultant residue by flash chromatography, elutingwith CH₂ Cl₂ :EtOAc (88:12). Combine the desired fractions and evaporatethe solvent. Dissolve the Cbz-diester (0.5 g) in ethanol (25 ml)containing 20% Pd(OH)₂ /C (0.15 g) and hydrogenate in a Parr apparatusat 50 p.s.i. at room temperature for 1 hour. Filter the resultantsolution through celite and evaporate the filtrate in vacuo.

Combine the resultant residue (0.33 g) and the product of Part C (0.59g) in dry DMF (7 ml), cool to 0° C. and add slowly1-hydroxybenzotriazole (0.18 g) followed by N-methyl morpholine (0.13g), then by DEC (0.25 g). Stir the mixture for 20 min. at 0° C., then atroom temperature for 16 hours or until TLC (silica, ethanol:methanol,85:15) indicates the reaction to be complete. Dilute the reactionmixture with CH₂ Cl₂ and extract with saturated NaHHCO₃, then with 10%aqueous citric acid. Dry the organic layer with MgSO₄, filter andevaporate the solvent in vacuo to obtain a residue.

E. Dissolve the product of Part D (0.75 g) in CH₂ Cl₂ (5 ml) and cool to0° C. Add, slowly and with stirring, trifluoroacetic acid (5 ml) andstir at 0° C. for 30 minutes, then at room temperature for 4 hours oruntil TLC (silica, CH₂ Cl₂ :MeOH, 90:10) indicates no starting materialis left. Evaporate the solvent.

Purify the resultant residue by ion exchange chromatography on BioradAG50-W-X2 resin (100-200 mesh, hydrogen form) previously equilibrated inethanol:water. Elute with ethanol:water:pyridine, combine the desiredfractions and evaporate the solvent to obtain a residue.

F. Treat the product of Part E in a manner similar to that described inExample 3, Parts F and G, substituting proline, t-butyl ester for theoctahydro-1H-indole to obtain the title compound.

EXAMPLE 8

N-[5-[[6-CHLORO-3,4-DIHYDRO-3-(2-PHENYLETHYL)-2H-1,2,4-BENZOTHIADIAZIN-7-YL]SULFONYLAMINO]-1-(S)-(METHOXYCARBONYL)PENTYN-3-YL]-(S)-ALANYL-(S)-PROLINE,S,S-DIOXIDE,HYDROCHLORIDE

A. Titrate a solution of diazomethane in ether into a solution of1-amino-1-(S)-carboxy-5-(t-butoxycarbonyl amino)-3-pentyne (4 g) inethanol (200 ml) until a yellow color remains. Evaporate the solvent toobtain 1-amino-1-(s)-methoxycarbonyl-5-(t-butoxycarbonylamino)-3pentyne.

B. Add the product of Part A (4 g) and PROTON SPONGE® (5 g) indichloromethane (15 ml) dropwise to a stirred solution of triflatereagent (6.5 g) (See Preparation 1) in dichloromethane (10 ml) at -10°C. Stir the solution and slowly allow to warm to room temperature over 2h. Filter the reaction mixture through celite, washing thoroughly withethyl acetate. Wash the combined organic solution with 10% citric acid(3×), saturated sodium bicarbonate (2×) and brine (2×), dry over MgSO₄and evaporate the solvent. Chromatograph on silica gel, eluting withethyl acetate:hexane (1:4) containing 1% triethylamine to obtain1-(S)-methoxycarbonyl-5-(t-butoxycarbonylamino)pentyn-3-yl-(S)-alanine,t-butyl ester.

C. Add the product of Part B (3.5 g) to a stirred solution of 4 M HCL indioxane (25 ml) at 0° and stir for 0.5 hours. Evaporate the solvent andtriturate the residue with ether to obtain1-(S)-methoxycarbonyl-5-amino-pentyn-3-yl-(S)-alanine, t-butyl ester,hydrochloride.

D. Add N-methylmorpholine (1.5 g) to a solution of the product of Part C(2.9 g) in tetrahydrofuran at 0°. Add6-chloro-3,4-dihydro-2-(phenylethyl)-2H-1,2,4-benzothiadiazine-7-sulfonylchloride, S, S-dioxide (3.9 g) (See Preparation 2) and stir theresulting mixture at room temperature overnight. Dilute the reactionmixture with ethyl acetate, wash with 0.5 N HCl (1×), saturated sodiumbicarbonate (2×), and brine (1×), dry over MgSO₄ and evaporate thesolvent. Chromatograph on silica gel, eluting with ethylacetate:dichloromethane to obtain a residue.

E. Add the product of Part D (4.8 g) to a 4 M solution of HCL in dioxane(100 ml) and stir the resulting mixture at room temperature overnight.Evaporate the solvent then triturate the residue with ether to obtain aresidue.

F. Treat the product of Part E in a manner similar to that described inExample 3, Part F, first paragraph, substituting (S)-proline, t-butylester for the octahydro-1H-indole, to obtain a residue.

Purify the resultant residue by chromatography on silica gel, elutingwith ethyl acetate:CH₂ Cl₂. Combine the desired fractions and evaporatethe solvent to obtain a residue.

G. Treat the product of Part F in a manner similar to that described inExample 3, Part G to obtain the title compound.

EXAMPLE 9N-[S-[[6-CHLORO-3,4,-DIHYDRO-3-(2-PHENYLETHYL)-2H-1,2,4-BENZOTHIADIAZIN-7-YL]SULFONYLAMINO]-1-(S)-METHOXYCARBONYL)-(E)-PENTEN-3-YL]-(S)-ALANYL-(S)-PROLINE,S, S-DIOXIDE,HYDROCHLORIDE

A. Add a solution of1-N-acetylamino-1-(S)-carboxy-5-(t-butoxycarbonylamino)-3-pentyne (5 g)in tetrahydrofuran (20 ml) dropwise to a stirred solution of sodium (1.2g) in liquid ammonia (1.2 L). After 2 h, add ammonium hydroxide andwater. Remove the ammonia, dissolve the residue in ethyl acetate, washwith 0.5 N HCl (2×) and brine (3×), dry over MgSO₄ and evaporate thesolvent. Purify by precipitation from dichloromethane as the DCHA saltof (E)-1-N-acetylamino-1-carboxy-5-(t-butoxy-carbonylamino)-3-pentene.

B. Add cobalt chloride hexahydrate (50 mg) and Acylase I (aminoacylasefrom porcine kidney, grade 1, available from Sigma Chemical Co., St.Louis, Mo.) (100 mg) to a stirred solution of the product of Part A (3.5g) in 0.1 M, pH 7.5 phosphate buffer (120 ml) at 38°. After 16 hours,remove the protein with activated carbon and filter through celite.Adjust pH of the solution to 2.75 and remove the unreacted isomer bywashing with ethyl acetate (3×). Adjust the pH to 6.5 and evaporate thesolvent. Remove the salt from the residue by dissolving in ethanol (100ml) and filtering through celite. Evaporate the solvent to obtain(E)-1-amino-1-(S)-carboxy-5-(t-butoxycarbonylamino)-3-pentene.

C. Substitute the product of Part B (i.e., the 3-pentene compound) forthe 3-pentyne compound in the procedure of Example 8, Parts B to G toobtain the title compound.

EXAMPLE 101-[2-(S)-[[2-[4-[[[6-CHLORO-3-CHLOROMETHYL-3,4-DIHYDRO-2H-1,2,4-BENZOTHIADIAZIN-7-YL]SULFONYLAMINO]METHYL]PHENYL-1-(S)-(ETHOXYCARBONYL)PROPYL]AMINO]-1-OXOPROPYL]-(2S-(2α,3aα,7aα)]-OCTAHYDRO-1-H-INDOLE-2-CARBOXYLICACID, T-BUTYL ESTER, S,S-DIOXIDE

A. To 1 g of the compound of Example 3, part B, in THF, addHCl-saturated dioxane (30 ml). Stir the mixture at room temperature for5 hours under an inert atmosphere. Evaporate the solvent to obtain a dryfoam as a hydrochloride salt.

B. Combine 3 g of the product of Part A with 2.5 g ofoctahydroindole-t-butyl ester (Preparation 3) in the manner described inExample 3, Part F to obtain1-[2(S)-[[p-cyanophenyl-1(S)-(ethoxycarbonyl)propyl]amino]-1-oxopropyl]-[2S-(2α,3aα,7aα)]-octahydro-1H-indole-2-carboxylicacid, t-butyl ester. FAB mass spec. M/e=501 (M⁺).

C. Hydrogenate 2.5 g of the product of Part B in 150 ml of ethyl alcoholcontaining 5 ml of saturated HCl/dioxane at 55 psi in the presence of500 mg of 10% Pd on C for 20 hours. Filter the resultant reactionmixture through celite to obtain 2.6 g of amine.

D. Combine 850 mg of the product of Part C with 600 mg of6-chloro-3-chloromethyl-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-sulfonylchloride, S,S-dioxide in 5 ml of THF containing 0.8 ml ofN-methylmorpholine and let stand overnight at room temperature. Dilutewith EtOAC, wash with saturated NaCl, and evaporate the solvent. Purifythe resultant residue by chromatography on SiO₂ with CH₂ Cl₂ /EtOAC aseluant to produce 700 mg of title compound. FAB mass spectrum showsM/e=844 (M⁺).

EXAMPLE 11N-[N-[4-[4-[[[[6-CHLORO-3,4-DIHYDRO-3-[2-(2-PYRIDINYL)ETHYL]-2H-1,2,4-BENZOTHIADIAZIN-7-YL]SULFONYLAMINO]METHYL]PHENYL]-1-(S)-ETHOXYCARBONYL)PROPYL]-(S)-ALANYL-(S)-PROLINE,T-BUTYL ESTER, S,S-DIOXIDE

A. To 6 g of the amine of Example 3, Part C, in 25 ml of DME cooled withan ice-bath, add 1.9 ml of N-methylmorpholine. To this mixture add 5 gof 4-amino-3-chloro-5-sulfonamidobenzenesulfonyl chloride in 25 ml ofDME. Stir the reaction overnight with cooling under an inert atmosphere.Concentrate the resultant reaction mixture and partition the residuebetween EtOAc and H₂ O. Separate the organic layer and evaporate thesolvent. Purify the resultant residue by chromatography on SiO₂ usingEtOAc/Hexanes as eluant to obtain 5 g of an oil. FAB mass spectrumM/e=633 (M⁺).

B. Treat the product of Part A in a manner similar to that described inExample 10, Part A, to yield a dry foam. FAB mass spectrum M/e=614 (M⁺).

C. Combine 1.6 g of the product of Part B with 490 mg L-proline, t-butylester in the presence of 9 ml of DMF, 380 mg 1-hydroxybenzotriazole,1.14 ml N-methylmorpholine and 550 mg DEC. Stir the reaction overnight,then extract and purify the resultant product as described in Example 3,Part F, using EtOAc/Hexanes as eluent to obtain a residue, FAB massspectrum M/e=730 M⁺.

D. In 7 ml THF, combine the amino-sulphonamide obtained in Part C with360 mg of (2-pyridinyl)propanal and 10 mg of p-toluenesulfonic acid andstir for 24 hours at room temperature in an inert atmosphere.Concentrate the reaction mixture and partition the resultant residuebetween EtOAc and water. Separate the organic phase, wash with diluteNaHSO₃, saturated NaHHCO₃, saturated NaCl and dry with MgSO₄. Evaporatethe solvent and chromatograph the residue on SiO₂ using EtOAc as eluantto obtain 350 mg of the title compound. Mass spectrum, M/e=899 (M⁺).

EXAMPLE 121-[N-[2-[4-[[[6-CHLORO-3,4-DIHYDRO-3-(2-PHENYLETHYL)-2H-1,2,4-BENZOTHIADIAZIN-7-YL]SULFONYL]AMINOMETHYL]-THIAZOL-2-YLMETHYLTHIO]-1(S)-(METHOXYCARBONYL)ETHYL]-(S)-ALANYL]-CIS,SYN-OCTAHYDROINDOLE-2(S)-CARBOXYLICACID, S,S-DIOXIDE (A) Ethyl-4-Aminomethylthiazole-2-Carboxylate 1.Ethyl-thizaole-4-Carboxylate

Combine ethylbromopyruvate (19.5 g, 100 mmole) in 50 ml EtOH andthioformamide (4.27 g, 70 mmole) in 10 ml EtOH and stir at roomtemperature overnight. Pour into 1 N HCl (100 ml) and extract with 190ml diethyl ether. Separate the aqueous layer and treat with an excess ofsolid sodium bicarbonate. Extract the aqueous layer with diethylether(2×150 ml), dry over MgSO₄ and evaporate the solvent to obtain the titlecompound of Part A(1) as a syrup. Recrystallize from hexanes to obtainwhite needles, m.p. 52°-54° C.

2. 2-Carboamide-Thiazole

Treat 5.0 g of the product of Part A(1) with 100 ml of concentratedammonium hydroxide solution overnight. Purge with N₂ for 2 hours andremove the solvent to obtain a crude product. Recrystallize usingethylacetate/pet ether to obtain the title compound, m.p. 122°-124° C.

3. 4-Cyano-Thiazole

Dissolve the product of Part A(2) (2.0 g, 14.50 mmoles) in 20 ml of dryCH₂ Cl₂ and cool in an ice bath. Add trifluoroacetic anhydride (3.65 g,16.50 mmoles) and stir at room temperature for 3 hours. Evaporate thesolvent and recrystallize from ethylacetate/pet ether to obtain thetitle compound, m.p. 54°-56° C.

4. Ethyl-4-Cyanothiazole-2-Carboxylate

Cool diisopropyl amine (4.04 g, 40.0 mmoles) in dry THF in an ice bathunder nitrogen. Add n-butyl lithium (40 ml, 40 mmoles) and stir at 0° C.for 15 min. Cool the resultant reaction mixture to -78° C. and add asolution of the product of part A(3) (4.0 g, 36.36 mmoles) in 20 ml THF.Stir the reaction mixture at -78° C. for 30 minutes, addethylchloroformate (3.93 g, 36.36 mmoles) and slowly warm the reactionmixture to room temperature. Add aqueous ammonium chloride and evaporatethe solvent. Add CH₂ Cl₂ (300 ml) to the resultant residue, wash withwater, dry over MgSO₄ and evaporate the solvent to obtain the titlecompound. Purify the product by column chromatography usingethylacetate/pet ether.

5. Ethyl-4-Aminomethylthiazole-2-Carboxylate

Cool a disiamylborane (17.50 mmoles) solution in an ice bath undernitrogen. Add the product of Part A(4) (1.5 g, 8.24 mmoles) in 10 ml ofdry THF and let stand at 0° C. for 2 days. Pour the reaction mixtureinto ice cold IN HCl and extract with diethyl ether. Separate theaqueous layer, treat with sodium bicarbonate, extract with 2×100 mlethyl acetate, dry over MgSO₄ and evaporate the solvent to obtain thetitle compound. Use without further purification in Part B.

B.1-N-[2-[4-[6-Chloro-3,4-Dihydro-3-(2-Phenylethyl)-2H-1,2,4,-Benzothiadiazin-7-yl]Sulfonyl]Aminomethyl]-2-Carboethoxythiazole,S,S-Dioxide

Combine the product of Preparation 2 (1.2 mmoles) and the product ofPart A (1.0 mmoles) in dry DMF (1.0 ml) and cool in an ice bath. Adddropwise triethyl amine (1.5 mmoles). Stir the resulting reactionmixture at room temperature for 16 hours. Evaporate the solvent underhigh vacuum, take up the residue in ethylacetate (100 ml), wash withwater, dry over MgSO₄ and evaporate the solvent. Purify the resultantresidue by column chromatography.

C.1-[N-[2-[4-[6-Chloro-3,4-Dihydro-3-(2-Phenylethyl)-2H-1,2,4-Benzothiadiazin-7-yl]Sulfonyl]Aminomethyl]-2-Hydroxymethylthiazole,S,S-Dioxide

Dissolve the product of Part B (1.0 mmole) in 25 ml dry THF and cool to-40° C. under nitrogen. Add LiALH₄ (1.5 mmole) and stir the resultantmixture at -40° C. for 1 hour. Add Na₂ SO₄ :10H₂ O, filter, dry overMgSO₄ and evaporate the solvent to obtain the title compound.

D.1-[N-[2-[4-[6-Chloro-3,4-Dihydro-3-(2-Phenylethyl)-2H-1,2,4-Benzothiadiazin-7-yl]Sulfonyl]Aminomethyl]-2-Bromomethylthiazole,S,S-Dioxide

Combine carbon tetrabromide (1.1 mmoles) and triphenyl phosphine (1.1mmoles) in dry DMF (5 ml) at 0° C. under nitrogen and stir at 0° C. for1/2 hour. Add a solution of the product of Part C (1.0 mmoles) in dryDMF (3 ml) and stir the reaction mixture at 0° C. for 1-2 hours. (Checkprogress of reaction by TLC). Pour the resultant mixture into ice coldwater, extract with diethyl ether (2×150 ml), dry over MgSO₄ andevaporate the solvent. Purify the resultant residue by columnchromatography.

E.Bis[[[1-(S)-(Methoxycarbonyl)-Ethyl]-(S)-Alanyl]-Cis,Syn-Octahydroindole-2(S)-tert-Butoxycarbonyl]-disulphide

1. Cool a solution of L-cysteine (7.20 g, 0.027 moles) in 50 ml dry CH₂Cl₂ in an ice bath under N₂. Add N-methylmorpholine (16.8 g, 0.16moles), then add a freshly prepared solution of triflate reagent (0.161moles) (Preparation 1) in 200 ml CH₂ Cl₂ dropwise over 2 hours. Stir theresulting reaction mixture overnight at room temperature. Wash thereaction mixture with 2×100 ml of water, then 2×100 ml of Columnchromatograph the resultant residue using 40% ethyl acetate:60% petether to obtain a syrup.

2. Cool a solution of the product of Part E(1) in 50 ml dioxane in anice bath. Add 150 ml of a solution of saturated dioxane:HCl and stir atroom temperature for 3 hours. Reduce the volume of the reaction mixtureto 50 ml and add 150 ml of ethyl acetate to precipitate the product asthe hydrochloride salt.

3. Cool a solution of the product of Part E(2) (1.0 mmole) in 10 ml dryDMF in an ice bath. Add 1-hydroxybenzotriazole hydrate (2.2 mmoles),1-(dimethylaminopropyl)-3-ethyl-carbodiimide (3.0 mmole) and2-carboxy-perhydroindole t-butyl ester (2.3 mmole) (Preparation 3) andstir the resulting solution at 0° C. Add triethylamine dropwise (6.0mmoles) and stir the resulting reaction mixture over-night at roomtemperature. Evaporate the solvent, dissolve the residue in 200 ml ofethylacetate, wash with water, brine, and dry over MgSO₄. Evaporate thesolvent and column chromatograph the resultant residue using 60%ethylacetate:40% pet ether.

F. Cool 5 ml of dry methanol in an ice bath under nitrogen, add sodiumborohydride (10.0 mmole) and stir at 0° C. for 10 minutes. Add theproduct of Part E (0.60 mmoles) in 5 ml dry methanol and stir theresulting mixture for 5 minutes at 0° C. Add the product of Part D (1.2mmoles) in 5 ml THF, warm the mixture to room temperature and stir atroom temperature for 2 hours. Evaporate the solvent and take up theresultant residue in ethylacetate. Extract with 10% citric acid, NaHHCO₃solution and brine, dry over MgSO₄ and evaporate the solvent.Chromatograph the residue to obtain the t-butyl ester of the titlecompound of Example 12.

G. Treat the product of Part F in a manner similar to that described inExample 3, Part G, to obtain the title compound. Mass spectrum m/e=925(M+).

EXAMPLE 131-[2(S)-[[4-[4-[[[6-CHLORO-3,4-DIHYDRO-3-(1-IMIDAZOLYLMETHYL)-2H-1,2,4-BENZOTHIADIAZIN-7-YL]SULFONYLAMINO]METHYL]PHENYL]-1(S)-(ETHOXYCARBONYL)BUTYL]AMINO]-1-OXOPROPYL]-CIS,SYN-OCTAHYDROINDOLE-2(S)-CARBOXYLICACID, DIHYDROCHLORIDE

A. Substituting 2-(4-cyano)phenylpropyl bromide for2-(4-cyano)phenylethyl bromide, carry out the procedure described inExample 3, Parts A-C to obtain the hydrochloride salt.

B. Treat the product of Part A as described in Example 11, Parts A andB, to obtain a residue.

C. Combine the product of Part B and perhydroindole-2-carboxylic acid,t-butyl ester (Preparation 3) as described in Example 11, part C andstir overnight. Evaporate the solvent and partition the resultantresidue between 5% NaHHCO₃ and EtOAc. Separate the EtOAc layer, dry overMgSO₄ and evaporate the solvent. Chromatograph the resultant residue byflash chromatography on SiO₂ (150 gm) eluting with 60% EtOAc: 40%hexane. Combine the desired fractions and evaporate the solvent toobtain a residue.

D. Dissolve sodium spheres (2.3 g) in dry methanol (100 ml) at roomtemperature, add imidazole (6.3 ml) and stir for 1 hour at roomtemperature. Add bromoacetaldehyde dimethylacetal (11.8 ml) and heat theresultant mixture overnight at reflux. Evaporate the solvent in vacuo,partition the resultant residue between EtOAc and water, dry the organiclayer over MgSO₄, and evaporate the solvent. Purify the resultantresidue by flash chromatography on SiO₂ (250 g), eluting with 4% CH₃ OHin CH₂ Cl₂. Combine the desired fractions and evaporate the solvent toobtain (1-imidazolyl)acetaldehyde dimethylacetal.

E. Combine the products of Part C (0.50 g) and Part D (0.13 g) in THF(20 ml), add HCl-saturated dioxane and stir, following the reaction bythin layer chromatography on silica gel (elute with 5% CH₃ OH in CH₂Cl₂) and adding HCl-dioxane until the reaction is complete. Evaporatethe solvent, partition the resultant residue between 5% NaHHCO₃ andEtOAc, wash the organic layer with brine, dry over MgSO₄, and evaporatethe solvent to obtain a residue. Purify the resultant residue by flashchromatography on SiO₂ (25 g), eluting with 3% CH₃ OH in CH₂ Cl₂.Combine the desired fractions and evaporate the solvent to obtain thet-butyl ester of the title compound.

F. Stir the product of Part E in HCl-saturated dioxane (200 ml) at roomtemperature and under nitrogen for 6 hours. Evaporate the solvent invacuo at room temperature to obtain the title compound, M/e=834 (M⁺).

In a similar manner, using appropriate starting materials and reagents,the following compounds may be prepared:

1-[2-(S)-[[1-(S)-carboxy-2-[4-[[[6-chloro-3-(cyclopentylmethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenylmethoxy]ethyl]amino]-1oxopropyl]-[2S-(2α,3aα, 7aα)]-octahydro-1H-indole-2-carboxylic acid, S,S-dioxide

1-[2-[[1-(S)-carboxy-2-[4-[[[6-chloro-3-(dichloromethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenylmethoxy]ethyl]amino]-1oxopropyl]-[2S-(2α,3aα, 7aα)]-octahydro-1H-indole-2-carboxylic acid, S,S-dioxide

1-[2-(S)-[[1-(S)-carboxy-3-[4-[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]propyl]amino]-1oxopropyl]-[2S-(2α,3aα, 7aα)]-octahydro-1H-indole-2-carboxylic acid, S,S-dioxide

1-[2-(S)-[[1-(S)-carboxy-3-[4-[[[6-chloro-3-(cyclopentylmethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]propyl]amino]-1-oxopropyl]-[2S-(2α,3aα, 7aα)]-octahydro-1H-indole-2-carboxylic acid, S,S-dioxide

1-[2-(S)-[[1-(S)-carboxy-3-[4-[[[6-chloro-3-(dichloromethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]propyl]amino]-1oxopropyl]-[2S-(2α,3aα, 7aα)]-octahydro-1H-indole-2-carboxylic acid, S,S-dioxide

1-[2-(S)-[[1-(S)-carboxy-2-[4-[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]ethyl]amino]-1-oxopropyl]-[2S-(2α,3aα, 7aα)]-octahydro-1H-indole-2-carboxylic acid, S,S-dioxide

N-[1-(S)-carboxy-3-[4-[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]propyl]-(S)-alanyl-(S)-proline,S,S-dioxide

N-[1-(S)-carboxy-3-[4-[[[6-chloro-3-(cyclopentylmethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]propyl]-(S)-alanyl-(S)proline,S,S-dioxide

N-[1-(S)-carboxy-3-[4-[[[6-chloro-3-(dichloromethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]propyl]-(S)-alanyl-(S)-proline,S,S-dioxide

N-[1-(S)-carboxy-2-[4-[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]ethyl]-(S)-alanyl-(S)-proline, S,S-dioxide

N-[1-(S)-carboxy-3-[4-[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]propyl]-(S)-alanyl-(S)-(4-cyclohexyl)proline,S,S-dioxide

7-N-[2-(S)-[[1-(S)-carboxy-3-[4-[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]propyl]amino]-1-oxopropyl]-1,4-dithia-7-azaspiro[4.4]nonane-8-(S)-carboxylicacid, S,S-dioxide

1-[2-(S)-[[1-(S)-carboxy-3-[4-[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]propyl]amino]-1-oxopropyl]-1H-2,3-dihydroindole-2-(S)-carboxylicacid S,S-dioxide

1-[2-(S)-[[2-[4-[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenylmethoxy]-1-(S)-[(2,2-dimethyl-1,3-dioxolan-4-yl)methoxycarboxyl]ethyl]amino]-1-oxopropyl]-[2S-(2α,3aα, 7aα)]-octahydro-1H-indole-2-carboxylic acid, S,S-dioxide

1-[2-(S)-[[3-[4-[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]-1-(S)-[(2,2-dimethyl-1,3-dioxolan-4-yl)methoxycarbonyl]propyl]amino]-1-oxopropyl]-[2S-(2α,3aα, 7aα)]-octahydro-1H-indole-2-carboxylic acid, S,S-dioxide

1-[2-(S)-[[3-[4-[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]-1-(S)-(ethoxycarbonyl)propyl]amino]-1-oxopropyl]-[2S-(2α,3aα, 7aα)]-octahydro-1H-indole-2-carboxylic acid, S,S-dioxide

N-[3-[4-[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]-1-(S)(ethoxycarbonyl)propyl]-(S)-alanyl-(S)-proline,S,S-dioxide

N-[3-[4-[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]-1-(S)-[(2-phenoxy)ethoxycarbonyl]propyl]-(S)-alanyl-(S)-proline,S,S-dioxide.

N-[3-[4-[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]-1-(S)-[(2,3-dihydroxy)propoxycarbonyl]propyl]-(S)-alanyl-(S)-proline,S,S-dioxide.

N-[3-[4-[[[6-chloro-3-(cyclopentylmethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]-1-(S)-(ethoxycarbonyl)propyl]-(S)-alanyl-(S)-proline,S,S-dioxide

N-[3-[4-[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]-1-(S)-[(2,2-dimethyl-1,3-dioxolan-4-yl)methoxycarbonyl]propyl]-(S)-alanyl-(S)-proline,S,S-dioxide

N-[3-[4-[[[6-chloro-3-(cyclopentylmethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]-1-(S)-[(2,2-dimethyl-1,3-dioxolan-4yl)methoxycarbonyl]propyl]-(S)-alanyl-(S)-proline,S,S-dioxide

N-[3-[4-[[[6-chloro-3-(cyclopentylmethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]-1-(S)-[(2-phenoxy)ethoxycarbonyl]propyl]-(S)-alanyl-(S)-proline,S,S-dioxide

N-[3-[4-[[[6-chloro-3-(cyclopentylmethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]-1-(S)-[(2,3-dihydroxy)propoxycarbonyl]-propyl]-(S)-alanyl-(S)-proline,S,S-dioxide

1-[2-[[2-[[4-[[[4-chloro-3-[[(phenylmethyl)amino]sulfonyl]phenyl]sulfonylamino]methyl]phenyl]methoxy]-1-(S)-(ethoxycarbonyl)ethyl]amino]-1-oxopropyl]-[2S(2α,3aα,7aα)]-octahydro-1H-indole-2-carboxylic acid

1-[2-[[2-[[4-[[[4-chloro-3-[[(phenylmethyl)amino]-sulfonyl]phenyl]sulfonylamino]methyl]phenyl]methoxy]-1-(S)-(carboxy)ethyl]amino]-1-oxopropyl]-[2S-(2α,3aα,7aα)]-octahydro-1H-indole-2-carboxylicacid

N-[2-[[4-[[[4-chloro-3-[[(phenylmethyl)amino]sulfonyl]phenyl]sulfonylamino]methyl]phenyl]methoxy]-1-(S)-(ethoxycarbonyl)ethyl]-(S)-alanyl-(S)-proline

N-[2-[[4-[[[4-chloro-3-[[(phenylmethyl)amino]-sulfonyl]-phenyl]sulfonylamino]methyl]phenyl]methoxy]-1-(S)-(carboxy)ethyl]-(S)-alanyl-(S)-proline

1-[2-[[2-[[4-[[[3-(amino)sulfonyl-4-chlorophenyl]-sulfonylamino]methyl]phenyl]methoxy]-1-(S)(ethoxycarbonyl)ethyl]amino]-1-oxopropyl]-[2S(2α,3a.alpha.,7aα)]-octahydro-1H-indole-2-carboxylicacid

1-[2-[[2-[[4-[[[3-(amino)sulfonyl-4-chlorophenyl]-sulfonylamino]methyl]phenyl]methoxy]-1-(S)-(carboxy)ethyl]amino]-1-oxopropyl]-[2S-(2α,3a.alpha.,7aα)]-octahydro-1H-indole-2-carboxylicacid

N-[2-[[4-[[[3-(amino)sulfonyl-4-chlorophenyl]-sulfonylamino]methyl]phenyl]methoxy]-1-(S)(ethoxycarbonyl)ethyl]-(S)-alanyl-(S)-proline

N-[2-[[4-[[[3-(amino)sulfonyl-4-chlorophenyl]sulfonylamino]methyl]phenyl]methoxy]-1-(S)(ethoxycarbonyl)ethyl]-(S)-lysyl-(S)-proline

N-[2-[[4-[[[3-(amino)sulfonyl-4-chlorophenyl]sulfonylamino]methyl]phenyl]methoxy]-1-(S)(carboxy)ethyl]-(S)-alanyl-(S)-proline

N-[2-[[4-[[[3-(amino)sulfonyl-4-chlorophenyl]sulfonylamino]methyl]phenyl]methoxy]-1-(S)-(carboxy)-ethyl]-(S)-lysyl-(S)-proline

N-[2-[[4-[[[3-(amino)sulfonyl-4-chlorophenyl]-sulfonylamino]methyl]phenyl]methoxy]-1-(S)-[(2,2-dimethyl-1-oxopropoxy)methoxycarbonyl]ethyl]-(S)-alanyl-(S)-proline

N-[3-[4-[[[6-chloro-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]-1-(S)(ethoxycarbonyl)propyl]-(S)-alanyl-(S)-proline,S, S-dioxide

N-[3-[4-[[[6-chloro-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]-1-(S)-[2,2-dimethyl-1-oxopropoxy)methoxycarbonyl]propyl]-(S)-alanyl-(S)-proline,S, S-dioxide

N-[3-[4-[[[6-chloro-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]-1-(S)(ethoxycarbonyl)propyl]-(S)-lysyl-(S)-proline,S, S-dioxide

N-[1-(S)-carboxy-3-[4-[[[6-chloro-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]-propyl]-(S)-alanyl-(S)-proline,S, S-dioxide

N-[1-(S)-carboxy-3-[4-[[[6-chloro-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]propyl]-(S)-lysyl-(S)-proline,S, S-dioxide

N-[1-(S)-carboxy-3-[4-[[[6-chloro-3-chloromethyl-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]-sulfonylamino]methyl]phenyl]propyl]-(S)-alanyl-(S)-proline,S, S-dioxide

N-[1-(S)-carboxy-3-[4-[[[6-chloro-3-chloromethyl-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]propyl]-(S)-lysyl-(S)-proline,S, S-dioxide

N-[3-[4-[[[6-chloro-3-chloromethyl-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]-1-(S)-[2,2-dimethyl-1-oxopropoxy)methoxycarbonyl]propyl]-(S)-alanyl-(S)-proline,S, S-dioxide

N-[3-[4-[[[6-chloro-3-chloromethyl-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]-1-(S)-(ethoxycarbonyl)propyl]-(S)-alanyl-(S)-proline,S, S-dioxide

1-[2-(S)-[[2-[4-[[[6-chloro-3-chloromethyl-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenylmethoxy]-1-(S)-(ethoxycarbonyl)ethyl]amino]-oxopropyl]-[2S-(2α,3aα,7aα)]-octahydro-1H-indole-2-carboxylicacid, S, S-dioxide

1-[2-(S)-[[1-(S)-carboxy-2-[4-[[[6-chloro-3-chloromethyl-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenyl]ethyl]amino]-1-oxopropyl]-2S-(2α,3aα,7aα)]-octahydro-1H-indole-2-carboxylicacid, S, S-dioxide

1-[2-(S)-[[1-(S)-carboxy-2-[4-[[[6-chloro-3-chloromethyl-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenylmethoxy]ethyl]amino]-1oxopropyl]-[2S-(2α,3aα,7aα)]-octahydro-1H-indole-2-carboxylicacid, S, S-dioxide

1-[2-(S)-[[1-(S)-carboxy-2-[4-[[[6-chloro-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenylmethoxy]ethyl]amino]-1-oxopropyl]-[2S-(2α,3aα,7aα)]-octahydro-1H-indole-2-carboxylicacid, S, S-dioxide

N-[1-(S)-carboxy-5-[6-chloro-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]pentyn-3-yl]-(S)-alanyl-(S)-proline,S, S-dioxide

N-[5-[6-chloro-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]-sulfonylamino]-l-(S)-(ethoxycarbonyl)pentyn-3-yl]-(S)-alanyl-(S)-proline,S, S-dioxide

1-[4-carboxy-5-[4-[[[6-chloro-3-(cyclopentylmethyl)-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-yl]sulfonylamino]methyl]phenylmethoxy]-1-oxopentyl]-octahydro-1H-indole-2-carboxylicacid, S, S-dioxide

N-[2-(S)-[1-(S)-carboxy-3-[4-[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonyl-amino]methyl]phenyl]propoxy]-1-oxopropyl]-(S)-proline,S, S-dioxide

N-[N-[3-[5-[[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonyl]amino]methyl]-1H-imidazol-2-yl]-1-(ethoxycarbonyl)propyl]-(S)-alanyl-(S)-proline,S,S-dioxide

N-[N-[3-[5-[[[[6-chloro-3,4-dihydro-3-(2-phenylethyl)-2H-1,2,4-benzothiadiazin-7-yl]sulfonyl]amino]methyl]-2-thiazolyl]-1-(ethoxycarbonyl)propyl]-(S)-alanyl-(S)-proline,S,S-dioxide

The following are non-limiting examples of a topical ophthalmicformulations of the invention. Compound A refers to1-[2-(S)-[[1-(S)-carboxy-5-[[6-chloro-3-chloromethyl-3,4-dihydro-2H-1,2,4-benzothiadiazin-7-y1]sulfonylamino]pentyl]amino]-1-oxopropyl]-[2S-(2α,3aα, 7aα)]-octahydro-1H-indole-2-carboxylic acid, S,S-dioxide. It iscontemplated, however, that therapeutically effective amounts of othercompounds of formula I may be substituted in its place.

EXAMPLE A

    ______________________________________                                        Topical Solution:                                                             Ingredients         mg/ml                                                     ______________________________________                                        Compound A           0.01                                                     Dibasic Sodium Phosphate                                                                          10.4                                                      Monobasic Sodium Phosphate                                                                        2.4                                                       Chlorobutanol       5.0                                                       Hydroxypropyl methylcelluose                                                                      5.0                                                       Sterile Water       q.s. ad 1.0 ml                                            1.ON NaOH           q.s. ad pH 7.4                                            ______________________________________                                    

Mix the ingredients under sterile conditions and using standardtechniques to obtain the ophthalmological solution.

    ______________________________________                                        Topical Solution:                                                             Ingredients             mg/ml                                                 ______________________________________                                        Compound A               0.01                                                 Timolol                 5.0                                                   Dibasic Sodium Phosphate                                                                              10.4                                                  Monobasic Sodium Phosphate                                                                            2.4                                                   Chlorobutanol           5.0                                                   Hydroxypropyl methylcelluose                                                                          5.0                                                   Sodium Hydroxide or Hydrochloric Acid                                                                 q.s. ad pH 7.4                                        Sterile Wter            q.s. ad 1.0 ml                                        ______________________________________                                    

Mix the ingredients under sterile conditions and using standardtechniques to obtain the ophthalmological solution.

Other beta adrenergic blocking agents may be used in the above examples,such as bulonol.

EXAMPLE C

    ______________________________________                                        Topical Solution:                                                             Ingredients             mg/ml                                                 ______________________________________                                        Compound A               0.01                                                 Dexamethasone Sodium Phosphate                                                                        1.0                                                   Dibasic Sodium Phosphate                                                                              10.4                                                  Monobasic Sodium Phosphate                                                                            2.4                                                   Chlorobutanol           5.0                                                   Hydroxypropyl methylcelluose                                                                          5.0                                                   Sodium Hydroxide or Hydrochloric Acid                                                                 q.s. ad pH 7.4                                        Sterile Wter            q.s. ad 1.0 ml                                        ______________________________________                                    

Mix the ingredients under sterile conditions and using standardtechniques to obtain the ophthalmological solution.

We claim:
 1. A method for reducing intraocular pressure which comprisestopically administering from about 0.000001% (w/v) to about 1.0% of anophthalmologically acceptable composition to the eye of a mammal in needof such treatment comprising a compound of the formula I: ##STR33## or apharmaceutically acceptable salt thereof, wherein: A is ##STR34## k is 1or 2; n is 0 or 1;p and q are 0, 1 or 2, provided that in structures IIband IIc the sum of p and q is 1 or 2, and that in formula IId, p is not0; B is --[J]--[L]--[M]--; D is ##STR35## E is --NH--, --O--, --S--, or--CH₂ --; G is --SO₂ --; J is --(CH₂)_(s) -- or --((CH₂)_(t) --W)--; Lis a chemical bond, cis- or trans-lower alkenylene, lower alkynylene,-Z-aryl-, -aryl-Z-, -Z-cycloalkyl-, or -cycloalkyl-Z-, a 5- or6-membered heterocyclic radical comprising 3-5 carbon atoms and 1 or 2heteroatoms selected from N, O and S, or a R⁵ -substituted heterocyclicradical, wherein aryl is ##STR36## and cycloalkyl is ##STR37## wherein wis 1, 2 or 3; M is --(CH₂)_(u) -- or --((CH₂)_(t) --X--(CH₂)_(v))--; Wis ##STR38## X and Z are independently a chemical bond, --NR⁹ --, --O--,--S--, ##STR39## s, u and v are independently 0-5; t is 1-5; R¹, R² andR⁹ are independently hydrogen, lower alkyl or lower acyl; R³ ishydrogen, lower alkyl, halo- and dihaloloweralkyl,trifluoroethylthiomethyl, phenylloweralkyl, (cycloalkyl)lower alkyl,aminomethyl, loweralkylaminomethyl, phenyl(lower)alkylthiomethyl,(cycloalkyl)loweralkylaminomethyl, 2-, 3- or 4-pyridyllower alkyl, 2-,4- or 5-, thiazolyloweralkyl, 2-, 4- or 5-1H-imidazolylloweralyl,1-imidazolylloweralkyl, 1-morpholinoloweralkyl orhydroxyphenylloweralkyl; R⁴ is chlorine or CF₃ ; R⁵ is hydrogen,halogen, lower alkyl, lower acyl, lower alkoxy, haloloweralkyl orphenylloweralkyl; R⁷ is hydrogen, lower alkyl or aminoloweralkyl; R⁶ andR⁸ are independently hydroxy, alkoxy having from 1 to 8 carbon atoms,benzyl, allyl, R¹⁰ --O_(r) --(CH₂)_(m) --O--, wherein O is oxygen orsulfur, r is 0 or 1 and m is 2 to 4, ##STR40## wherein the alkyl hasfrom 3 to 8 carbon atoms, ##STR41## wherein the phenyl may besubstituted with group T defined below, 1-glyceryl, ##STR42## R¹⁰ isphenyl, substituted phenyl wherein the substituents are chosen fromgroup T, 1-naphthyl or 2-naphthyl; T is halogen, hydroxy,trifluoromethyl, lower alkoxy, lower alkyl, 2-furanyl, 3-furanyl,2-thienyl, 3-thienyl, phenyl and substituted phenyl wherein thesubstituents are chosen from halogen, hydroxy, trifluoromethyl, loweralkoxy or lower alkyl; R¹¹ is hydrogen or alkyl having from 1 to 8carbon atoms; R¹² is hydrogen, lower alkyl, unsubstituted or substitutedphenyl and substituted or unsubstituted phenyl lower alkyl, whereinphenyl may be substituted by group T; and R¹³ is hydrogen or loweralkyl; provided that if L is alkenylene or alkynylene, J is --(CH₂)_(s)-- wherein s is 1-5; provided that if L is -Z-aryl-or -Z-cycloalkyl-, Jis --(CH₂)_(s) -- wherein s is 2-5; provided that if L is alkenylene,alkynylene; -aryl-Z- or -cycloalkyl-Z-, M is --(CH₂)_(u) -- wherein u is1-5; provided that if s and u are each zero, L is aryl or cycloalkyl(i.e. Z is a bond); and provided that if s and v are each zero, L isaryl or cycloalkyl (i.e. Z is a bond)in combination with from about 0.01to about 1.0% of a beta adrenergic blocking agent.
 2. A method accordingto claim 1 wherein the beta adrenergic blocking agent and the compoundof formula I are administered topically together in the samecomposition.
 3. A method according to claim 2 wherein the betaadrenergic blocking agent is selected from atenolol, metoprolol,nadolol, pindolol, propranolol, timolol, labetalol, cartelol, bunolol,betaxolol or dilevalol or pharmaceutically acceptable salts and/orisomers thereof.
 4. A method according to claim 2 wherein the betaadrenergic blocking agent is timolol.
 5. A method according to claim 2wherein the beta adrenergic blocking agent is bunolol.
 6. A topicalophthalmologically acceptable composition useful for controllingelevated intraocular pressure which comprises from about 0.01 to about1.0% of a beta adrenergic blocking agent, and from about 0.000001 toabout 1.0% of a compound represented by the formula I: ##STR43## or apharmaceutically acceptable salt thereof, in combination with anophthalmologically acceptable carrier for topical use, wherein:A is##STR44## k is 1 or 2; n is 0 or 1; p and q are 0, 1 or 2, provided thatin structures IIb and IIc the sum of p and q is 1 or 2, and that informula IId, p is not 0; B is --[J]--[L]--[M]--; D is ##STR45## E is--NH--, --O--, --S--, or --CH₂ --; G is --SO₂ -- J is --(CH₂)_(s) -- or--((CH₂)_(t) --W)--; L is a chemical bond, cis- or trans-loweralkenylene, lower alkynylene, -Z-aryl-, -aryl-Z-, -Z-cycloalkyl-, or-cycloalkyl-Z-, a 5- or 6-membered heterocyclic radical comprising 3-5carbon atoms and 1 or 2 heteroatoms selected from N, O and S, or a R⁵-substituted heterocyclic radical, wherein aryl is ##STR46## andcycloalkyl is ##STR47## wherein w is 1, 2 or 3; M is --(CH₂)_(u) -- or--((CH₂)_(t) --X--(CH₂)_(v))--; W is ##STR48## X and Z are independentlya chemical bond, --NR⁹ --, --O--, --S--, ##STR49## s, u and v areindependently 0-5; t is 1-5; R¹, R² and R⁹ are independently hydrogen,lower alkyl or lower acyl; R³ is hydrogen, lower alkyl, halo- anddihaloloweralkyl, trifluoroethylthiomethyl, phenylloweralkyl,(cycloalkyl)lower alkyl, aminomethyl, loweralkylaminomethyl,phenyl(lower)alkylthiomethyl, (cycloalkyl)loweralkylaminomethyl, 2-, 3-or 4-pyridylloweralkyl, 2-, 4- or 5-, thiazolyloweralkyl, 2-, 4- or5-1H-imidazolylloweralyl, 1-imidazolylloweralkyl, 1-morpholinoloweralkylor hydroxyphenylloweralkyl; R⁴ is chlorine or CF₃ ; R⁵ is hydrogen,halogen, lower alkyl, lower acyl, lower alkoxy, haloloweralkyl orphenylloweralkyl; R⁷ is hydrogen, lower alkyl or aminoloweralkyl; R⁶ andR⁸ are independently hydroxy, alkoxy having from 1 to 8 carbon atoms,benzyl, allyl, R¹⁰ --O_(r) --(CH₂)_(m) --O--, wherein O is oxygen orsulfur, r is 0 or 1 and m is 2 to 4, ##STR50## wherein the alkyl hasfrom 3 to 8 carbon atoms, ##STR51## wherein the phenyl may besubstituted with group T defined below, 1-glyceryl, ##STR52## R¹⁰ isphenyl, substituted phenyl wherein the substituents are chosen fromgroup T, 1-naphthyl or 2-naphthyl; T is halogen, hydroxy,trifluoromethyl, lower alkoxy, lower alkyl, 2-furanyl, 3-furanyl,2-thienyl, 3-thienyl, phenyl and substituted phenyl wherein thesubstituents are chosen from halogen, hydroxy, trifluoromethyl, loweralkoxy or lower alkyl; R¹¹ is hydrogen or alkyl having from 1 to 8carbon atoms; R¹² is hydrogen, lower alkyl, unsubstituted or substitutedphenyl and substituted or unsubstituted phenyl lower alkyl, whereinphenyl may be substituted by group T; and R¹³ is hydrogen or loweralkyl; provided that if L is alkenylene or alkynylene, J is --(CH₂)_(s)-- wherein s is 1-5; provided that if L is -Z-aryl-or -Z-cycloalkyl-, Jis --(CH₂)_(s) -- wherein s is 2-5; provided that if L is alkenylene,alkynylene; -aryl-Z- or -cycloalkyl-Z-, M is --(CH₂)_(u) -- wherein u is1-5; provided that if s and u are each zero, L is aryl or cycloalkyl(i.e. Z is a bond); and provided that if s and v are each zero, L isaryl or cycloalkyl (i.e. Z is a bond).
 7. A composition according toclaim 6 wherein said beta adrenergic blocking agent is selected fromatenolol, metoprolol, nadolol, pindolol, propranolol, timolol,labetalol, betaxolol, carteolol, bunolol or dilevatlol, orpharmaceutically acceptable salts and/or isomers thereof.
 8. Acomposition according to claim 6 wherein said beta adrenergic blockingagent is timolol.
 9. A composition according to claim 6 wherein saidbeta adrenergic blocking agent is bunolol.